Good morning, class.
Nice to see you again.
Hope you had a great weekend.
If you didn't, it wasn't because of the weather.
So here I am, once again a member of the walking wounded,
and we're talking about carbohydrates today,
as you may recall, or at least we
were at the end of our discussion last time.
And, we made the point that these multiple hydroxyl groups
on the carbohydrates, on the one hand,
determine the identity of various kinds of sugars.
Just the orientation, the three-dimensional orientation
of them, for one thing, and for another
that these multiple hydroxyl groups represent
the opportunity for forming covalent bonds
with other monosaccharides as is indicated here
in these disaccharides, or covalent bonds end-to-end
to create large molecules, which will increasingly
be the theme of our discussion today, i.e.
when I talk about large molecules,
we just used the phrase generically, macromolecules,
since in principle these end to end joinings of molecules which
involve the dehydration and the formation
of these covalent bonds like right here
can create molecules that are hundreds, indeed even
thousands of subunits long.
So, here, if we're talking about a polymer,
we refer to each one of these subunits
of the polymer as being a monomer,
and the aggregate as a whole as being a polymer.
Here, we touched upon the fact toward the end of last lecture,
in fact, at the very end, that one can cross-link
these long, linear chains of carbohydrates.
And here, we see the fact that glycogen,
which is a form of glucose that is stored in our liver
largely, and to a small extent in the muscles
actually is cross-branched.
So, if one draws on a much smaller scale a glycogen
molecule, one might draw a picture that looks like this.
And it looks almost like a Christmas tree
with multiple branches.
And, the purpose of this is actually
to sequester the glucose, to store the glucose
into metabolically inactive form until the time comes
that the organism needs, once again,
the energy that is stored in the glucose
upon which occasion these bonds are rapidly broken down
and the glucose is mobilized and put
into the circulation for eventual disposition
and use in certain, specific tissues.
While it's encumbered in these high molecular weight polymers,
the glucose is essentially metabolically inactive.
The body doesn't realize it's there.
And, we can, as a consequence store large amounts of energy
in these glycogen molecules.
And it can be stored there indefinitely.
Now, the fact is this idea of end-to-end polymerization
that I just mentioned can be extended
to other macromolecules which also become linked end-to-end
in specific kinds of polymers.
And here, we are moving, now, into the notion
of talking about amino acids.
And, we're talking about proteins.
If we look at an amino acid, what we see
is it has an important structure like this.
Here's a central carbon ?
for, in principle, the distinct side
chains where R represents some side chain that
can be any one of, as we'll see shortly,
20 distinct identities.
But, all the amino acids share in common the property
that they have this overall structure.
And, as you may recall from our discussions of last week,
at neutral pH, an amino acid of this sort, whatever
R is, wouldn't look like this at all
because the amine group would attract an extra proton,
causing it to become positively charged.
And, the carboxyl group would release a proton,
causing it to become negatively charged.
And, as you might deduce from this,
at very low pHn due to the greatly increased concentration
of protons, free protons in the solution,
the equilibrium would be driven more in favor
of reattaching a proton to the carboxyl group
just because there are so many of these protons around.
Conversely, at very high pH, where the hydroxyl ions are
in predominance, they obviously tend to scavenge protons,
reducing the level of protons to very low levels in the water.
And, under very high pH conditions,
this proton would be released and pulled away
by the hydroxyl ions causing this amine group once again
to return to its negative charge state.
Now, the fact of the matter is that these amino acids
exist in a very specific three-dimensional
configuration.
And that's illustrated much more nicely here
than I could possibly draw on the board, which in any case
would be hopeless.
And, you can see the principle that once
you have four distinct side groups coming off of carbon,
that there is, in principle, two different ways to create them.
And, this is sometimes called chirality.
Chiral, you see, is the form right here.
The hands are chiral.
If I try, as much as I will, to superimpose
one hand on top of another.
It doesn't work because they are mirror images of one another,
which are asymmetrical.
And, as a consequence, we see a similar kind of relationship
occurring here where we see that these two forms of amino acids
could, in principle, exist.
And, they are not interchangeable
unless one breaks one of the bonds and reforms it.
These two forms are called the L and the D,
and it turns out that the L form is the one that's
used by virtually all life forms on the planet, i.e.
there was an arbitrary choice made
sometime about 3 billion years ago or more to use
one of the three dimensional configurations,
and not to use the other.
The other is found in certain rare exceptions,
but virtually all life forms on this planet use the L form.
That said, by the way, this indicates some of the arbitrary
decisions that were made early during evolution because we
could imagine on another planet if life were to exist there
and it were to depend on amino acids,
and that evolutionary system might have chosen the D form.
So, this is sort of a luck of the draw.
This is actually the way things evolved here.
And, what we begin to see, now, is if we talk about proteins
or if we wanted to be more specific and use the more
biochemical term ?polypeptide,?
we see once again we have an end-to-end joining system which
is a bit different from that which
the monosaccharides employ to create long chains of glycogen
or of starch because here we see once again a dehydration
reaction where an amine group and a carboxyl group
are caused to shed their hydroxyl and the proton,
causing the formation of a peptide bond.
And here, we see this important, very important
biochemical entity, a peptide bond consisting here
of this carbonyl and this nitrogen fused
in this specific way.
And, of course, if you recognize this as being a peptide bond,
then you can understand why proteins are sometimes given
the term ?polypeptide.?
In some cases, if one has very short stretches of amino acids
linked end to end like this, we talk about these being
oligopeptides, where ?oligo?
is the general term used in biology
to refer to a small number of things
rather than a large number of things.
And, once again, we have, here, the possibility
of extending this infinitely.
There are no constraints, in principle,
on making this 500, 1,000, even 2,000 amino acids long,
where each one of these, once again, is an amino acid,
and where once again I'm being very coy about the identities
of R1 and R2, which, as I will indicate very shortly,
can be one of 20 distinct alternatives.
Here, you see that we are continuing
this process of peptide bond formation.
And most importantly here is the realization
that there is a polarity of elongation here.
It doesn't move with equal probability left or right,
or right to left.
We start at the amino end here.
This is the amino end, and this is the carboxyl end.
The amino end and the carboxyl end,
and invariably, again because of the way life
has evolved on this planet, the new amino acid
is added on the carboxyl end.
And so, when one often talks about proteins,
one refers to their N terminal, and to their C terminal
ends, these referring obviously to the amino group
at one end and a carboxyl and at the other end
so that polarity is always a directed synthesis adding it
on C-terminal end, in other words
to use a short-hand notation, we think about proteins
as going with this polarity N toward C.
Things are growing at the C terminal end progressively.
And, each time one can imagine the addition of an amino acid
on the end of it.
So, again, it can be extended, in principle, indefinitely.
Keep in mind as well, something that's implicit in everything
I'm telling you but I won't always mention it explicitly,
and that is virtually every biochemical reaction
is reversible.
And therefore, if one is able to form a peptide bond,
one is able to break it down by biological means as well, i.e.
by introducing a water molecule back in and thereby using
the process of hydrolysis, which is the breakdown
of a bond through the introduction of a water
molecule to destroy the previously created bond.
To use an MIT phrase, the reversibility is intuitively
obvious because if you are able to make a, well,
I don't know if it?s still used, but it was used in the late
Stone Age around here.
Anyhow, any biochemical action must be reversible
because if, for example, this polymerization were
irreversible, then all the protein that was ever
synthesized on the surface of the planet over the last 3 1/2
billion years would accumulate progressively.
And obviously, that doesn't happen.
And therefore, macromolecular synthesis,
to the extent that it proceeds forward,
obviously must go the other direction as well.
And the resulting concentration of a complete protein
is known as its steady state.
So we might make a protein at one rate
and break it down at the same rate.
And its steady-state concentration
represents the compromise between these two, i.e.,
the concentration of such a protein
that we might observe at any one point in time.
Indeed, the term ?steady-state?
could be expanded to any process in which there is a synthesis
and there is a breakdown of something.
And the equilibrium concentration
which results is, once again, called the steady-state
of that molecule.
Now, let's get down to the nitty-gritty,
which is obviously something which
we can't avoid for very long, which is to say the R's, i.e.
the side chains.
Once again, here we see an arbitrary artifact
of very early evolution in the biosphere
because there are, in effect, 20 different side chains creating
20 distinct amino acids, which are
used in proteins by all organisms on this planet.
Again, there are rare exceptions,
certain fungi and certain bacteria
are able to make unusual amino acids.
But these are the basic building blocks
of virtually all life forms on the planet.
99.99% of all the protein that is created
is synthesized through the polymerization of these 20
amino acids.
And, by the way, one of the amino acids, glycine,
over here, you see it right here,
violates this rule of chirality.
And, you will recall before I said
that because there are four distinct amino acids,
four distinct side chains around a central carbon
sometimes called the alpha carbon,
you always have a handedness of amino acids.
But this notion cannot be respected in the case
of glycine seen up here simply because we don't have four
distinct, here's the central carbon where I'm pointing with
the red, and here these two hydrogens are equivalent to one
another.
They are not four distinct chains.
There's only three distinct chains here.
So glycine violates this rule of chirality,
of left- and right-handedness.
And here, by the way, the side chain,
which in all of these cases is depicted as extending off
to the right of each amino acid, the side chain
is simply an H, simply a proton, a hydrogen atom.
In fact, what we see about these amino acids
is that the side chains have quite
distinct biochemical properties.
And that begins to impress us with the notion
that proteins and their biochemical attributes
can be dictated by the identities of the amino acids
that are used to construct them.
We can talk about the notion of nonpolar
versus polar amino acids, i.e., amino acids
which have poor affinity for water.
They don't have a separation of plus and minus charges.
And as a consequence, they are a little bit or quite a bit
hydrophobic.
Now, you will say, well, how can they be hydrophobic,
because here this oxygen is charged,
and here this amine group is charged?
That would make it highly hydrophilic.
But keep in mind, when I'm talking
about these amino acids, I'm not talking about them
when they are in a single amino acid form.
I'm talking about their properties
once they have been polymerized into state like this.
And, once they are polymerized into state like this,
the NH2 and CO charging, that is, the charge here
and the charge here become irrelevant
because this oxygen and this amine group
are both tied up in covalent bonds.
And, this acquisition of a proton and this shedding
of a proton over here cannot occur,
because both of these atoms, O and N,
are involved in covalent bonds.
So therefore, when we talk about nonpolar and polar amino acids,
keep in mind we are focusing on the biochemical properties
of the side chain because the central backbone
of the polypeptide and the central backbone
is defined quite clearly here.
Here's the central backbone, and you
see it has a quite repeating structure, N, C, C, N, C, C, N,
C, C, this is invariant.
What changes, and what defines the biochemical attributes
of this oligopeptide, or a polypeptide,
are the identities of these side chains, which
again are plotted on this particular graph.
You have a different version in your book off to the right.
Here, you see, we have a proton, a methyl group, a valine,
a lucine, an isolucine, and the differences
between this suggests these are all quite aliphatic, quite
similar to the propane that we talked about last time,
or the hexane.
That is to say, these are quite hydrophobic side groups.
And, as such, if there were a polypeptide, we can imagine,
and you put the polypeptide in water,
you can imagine that these amino acids would not
like to be directly confronting the water because of the fact
that they are hydrophobic.
Methionine is also a bit hydrophobic.
I'm equivocating there because the S
has a slight degree of hydrophilicity.
It has a slight degree of polarity,
but not really that much.
And, these aromatic side chains here,
because they have these benzene rings,
consequently are called aromatic.
These are quite strongly hydrophobic.
So, they really hate to be in the intimate contact
with water.
Here, on the other hand, let's look at these side chains
because here we have strongly polar molecules,
side chains again.
Keep in mind we are focusing on the side chains.
Here we see serine with the hydroxyl group that
can form hydrogen bonds with the water, threonine, which
has its own hydroxyl group, asparagine,
which has two atoms here, this carbonyl and the NH2,
both of which can form hydrogen bonds with the water,
as can glutamine.
So, these are quite hydrophilic.
They are not as fanatically hydrophilic as these charge
molecules where the side chains are not
just capable of forming hydrogen bonds.
In this lower group here, the side chains
are capable of undergoing ionization.
So they're actually strongly charged.
And here, we see here the carboxyl group,
and our aspartic acid and glutamic acid
has actually discharged its proton,
becoming negatively charged.
These are acidic amino acids, by virtue
of the carboxyl group they have, basic amino acids
here: arginine, lysine, and histidine,
all acquire a positively charged side
chain by virtue of these nitrogen
here which have a strong affinity for pulling away
protons or abstracting protons from the aqueous solvent.
And so, we have a whole gradient of hydrophilicity
down to hydrophobicity.
And here, we have intermediate structures.
We also have some very special idiosyncratic kinds
of amino acids.
Here is tyrosine, and tyrosine is little bit schizophrenic
again.
It has this highly hydrophobic aromatic group here,
the benzene ring, which hates to be in water,
and the hydroxyl group which actually is a friend of water.
So, here, we have something where
its role is quite equivocal.
Here, we have cystine, as indicated here,
and what's interesting about the cystine group
in this case is the SH group, the side chain, the SH
group, because this SH group is able to form bonds with yet
other SH groups from other cystines.
So, let's just look at the cystine here for a moment.
You see there's a CH2, and then there's an SH.
So, let's imagine, I'm not going to draw all the atoms here,
but let's imagine here we have the CH2 group.
I'm not drawing the backbone, SH, over here.
And, we can imagine another protein chain,
another polypeptide chain down here.
Again, I'm not drawing the backbone,
but I'm drawing another SH like this.
And, the fact is, under the conditions of oxidation
and reduction that operate at least in the extracellular
space, one can oxidize this, these two,
resulting in the formation of what
is known as a disulfide bond.
So, here, we have now for the first notion
the idea that the polypeptide chains can be covalently linked
to one another through these cross-links, as indicated here.
Conversely, if you add a reducing agent
that will add protons back to this,
and reduce the oxidation state of the sulfurs,
once again causing the disulfide bond to fall apart.
Now, in principle, these disulfide bonds
could be used to link two proteins together.
But, more often than not, if you look
at the structure of a single protein,
here's the structure of a single protein.
And often, there are intramolecular bonds,
disulfide bonds, i.e., bonds from one domain
of the protein to another, from one part of the protein
to the other.
I'll draw them in right here.
Here might be a disulfide bond.
Here might be a disulfide bond, and I could go on and on.
There might be another one over here.
Why do we have these disulfide bonds?
Because as we will indicate very shortly,
the three-dimensional structure of a protein
is very specifically determined.
A protein can only function when it
assumes a certain three-dimensional
configuration, when it assumes a certain three-dimensional,
stereochemical configuration.
When we talk about stereochemistry,
we are talking about the three-dimensional structures
of molecules, small and large.
And, here, we begin to touch on a theme of how
these complex polypeptide chains are able to create proteins
that have very specific, often very rigid,
structures in three-dimensional space.
Part of this structural rigidity is maintained
by these covalent disulfide bonds, which
tightly link neighboring regions, or even not
so neighboring regions, of a single polypeptide chain,
these intramolecular links.
This doesn't preclude there being intermolecular links
between two polypeptide chains that
are mediated as well by the disulfide bonds.
Here's another very peculiar amino acid
because what you see here is at the side chain, which
is CH2, CH2, CH2, CH2 is hydrogen bonded here
to the amine group.
It's not swinging out in free space.
I misspoke.
What we see here is CH2, CH2 is covalent bonded
to the amine group.
You pick that up, right?
I was just testing you.
Sure I was.
OK, so here we see a five-membered ring
that's created.
So here, this thing is not swinging out in free space.
It creates a five-membered ring where the end of the side chain
is actually covalently linked to the amino group.
And, that also has implications for the structure of proteins
because this particular amino acid, whenever
it occurs within a polypeptide chain,
doesn't have the flexibility of assuming certain configurations
that the other ones have whose four side chains are not
so encumbered.
None of them has total flexibility,
but this one is far more encumbered
in the kinds of three-dimensional structures
that it can assume.
And, with that in mind, we begin to ask questions
about how polypeptide chains assume
three-dimensional structure.
If we talk about a polypeptide chain, in our minds,
hopefully, there's only 28 combinations.
Oh, am I good or what?
Anyhow, all right, so, look here.
And here, you see, this is a typical polypeptide chain.
Here, we have a three letter code.
In truth, there is a single letter code
which was introduced around 1965.
So, each of the 20 amino acids has its own single letter code.
And, to make a frank and depressing admission, 35 years,
40 years after the single amino acid letter
code was instituted, I still haven't learned it.
But, we could learn these three letter codes,
which fortunately are present here.
In the single letter code L is lucine,
and A is alanine, see, they know it.
This is another example of not being
able to teach old dogs new tricks.
Anyhow, so here we see the way, one way
by which one might depict an amino acid chain,
a polypeptide chain.
And keep in mind, this can go on indefinitely.
As we begin to wrestle with the three-dimensional structure
of the chain, we begin to realize the following,
and that is that after the chain is initially synthesized,
it's initially chaotic.
And, as it extends, it increasingly
begins to assume a very specific three-dimensional molecular
configuration which is indicated down here.
So, the chaos that operates initially
will eventually result in a native configuration over here,
which in many respects often represents the lowest
free energy state.
Since for the last 40 years, people have been trying
to figure out, if you knew the amino acid sequence of this
primary polypeptide here, if you knew its primary structure,
and when I say, ?primary structure,?
what I mean is the sequence of the amino acids.
So, if you knew the primary structure of the amino acids,
you should, in principle, be able to develop a computer
algorithm that would predict the three-dimensional
configuration, which is shown here in a very schematic way,
and which we will discuss in much greater detail shortly.
And, the fact is, after 40 years of trying,
one still is unable to do that, i.e.,
if I were to give the primary amino acid
sequence of the polypeptide to the smartest
biochemists in the world, and there are some very smart ones,
he or she could still not tell me
what the three-dimensional structure
of this protein with total certainty would be.
Why?
Because there's an almost infinite number
of intramolecular interactions that greatly
complicate how the protein assumes the structure.
Moreover, if we talk about this as the native state
of the protein, we can imagine that there?s ways of disrupting
that because much of this native state is created
by intramolecular hydrogen bonds.
Remember, the hydrogen bonds are relatively weak,
and if we heat up the temperature,
then we can break hydrogen bonds.
And therefore, every time we fry an egg, for example,
if we want to get down to Earth, we denature.
We break up the native three-dimensional structure
of the albumin molecules that constitute the egg white.
And so, when everything turns white, what we've done
is to take a native molecule like this, heated it
up to temperatures where the intramolecular
bonds no longer stabilize.
Notably, hydrogen bonds no longer
stabilize this three-dimensional configuration,
and we put it into a denatured state, which
might be all the way up here.
And, therefore, this acquisition of a native configuration,
or a native state, native representing the natural state,
is also reversible in many molecules
simply by heating them up.
There are to be sure yet other molecules which
are different from the egg white from the albumin in egg
white where if you cool them back down,
they will spontaneously reassume their native structure.
Many proteins, most, will not do so.
Well, again, let's go back to this issue of the acquisition
of complex, three-dimensional structure.
And here, we begin to see how some of this structure
is acquired and stabilized through these intramolecular
hydrogen bonds.
And there are many opportunities for these intramolecular
hydrogen bonds because here we see one polypeptide chain here,
here we see another.
And, we see that the NH2 group right here, I'm sorry,
the nitrogen group here with the proton side chain,
and the carbonyl group here with the oxygen are not encumbered.
They are, in principle, available to form hydrogen
bonds with a polypeptide chain somewhere else.
Now, this other polypeptide chain
could once again be from another protein,
from another polypeptide.
But more often than not, we are once again
dealing with intramolecular cross-links.
But in this case, the intramolecular cross-links
are not disulfide bonds which are covalent,
and hard and stable as a rock in the absence of reducing agents.
Here, we're talking about much weaker bonds, hydrogen bonds
which also act between different loops of the protein
and serve, once again, to stabilize
the three-dimensional structure, the native state
of the protein.
And, you can see how these opportunities for forming
multiple hydrogen bonds can create
an enormous degree of stability.
And, here are some examples of what we now
call the secondary structure of the protein.
Just a second ago, or several minutes ago to be honest,
and I'm always honest with you, class,
the primary structure is the amino acid sequence.
The secondary structure represents
configurations like this.
Here is an alpha helix.
Here is a beta pleated sheet.
And, what we see here in this alpha helix
is we have a helical structure where the amine group down
here, the NH group, hydrogen bonds with a residue that is,
I think, three and a half residues upstream,
one, two, there's an amine down there, so,
with the carbonyl group that's three and a half residues
upstream of it.
This one, once again, reaches three and a half residues
upstream.
Not all the hydrogen bonds are shown in the background.
Only the ones on our side of the helix
are shown, on the front side of the helix.
But, you can imagine that this can perpetuate itself.
And, each of these carbonyl's may
associate with a proton, an NH group that's
either above or below that particular residue.
And this, in turn, can create a helical structure.
By the way, proline doesn't fit well.
If you add a proline in here, proline
is known in the trade as a helix breaker.
Why?
Because it cannot twist itself around to form an alpha helix.
And so, if the primary amino acid
were to dictate that a proline would be inserted right
here, for example, then this helix
might exist down below and above,
but it would not be continuous because the presence
of a proline is highly disruptive of the formation
of an alpha helix.
This means that, in principle, you
can make some predictions about the localized
structure of a polypeptide by knowing whether or not
proline is present, for example.
But that still doesn't give you the power
to predict the entire three-dimensional structure
of the finished protein itself.
Now, let's agree that this is the secondary structure
of the protein, i.e., the various domains which often
form alpha helices within a certain segment of the protein
or a certain segment of the protein
will form beta pleated sheets.
And there are several other less common kinds
of secondary structure.
And here, we deal with tertiary structure.
Now we are getting really interesting.
Or, maybe you don't like it.
But some people say it's really interesting
because here are the tertiary structures of some arbitrarily
chosen proteins.
Here, the tertiary structure of this particular protein,
and the identities of these are not given in our textbook.
And, I'm sure if we spent two or three weeks,
we could find out what they were.
But anyhow, here is a protein, a three-dimensional structure
of a protein which is composed of four alpha helices which
go up.
Another alpha helix, alpha helix, alpha helix, alpha
helix, they are depicted here, fortunately,
in four different colors.
And so, we see that what we talk about tertiary structure,
we're talking about how the alpha helices are disposed
with respect to one another.
The primary structure of the amino acid sequence
is not shown here.
The secondary structure represents these individual
alpha helices, and the tertiary structure represents how these
alpha helices are arranged vis-?-vis one another.
Here is a protein which is structured much differently.
It's formed of many beta pleated sheets.
We saw that in the last figure, in the last overhead.
You see it as a quite different overall three-dimensional
structure.
This could be the beginning of an alpha helix down here,
although that's quite equivocal.
And here, we see yet another point.
And that is, as we said before, the tertiary structure
independent of these alpha helices and beta pleated sheets
may be stabilized by these covalent inter-strand
cross-links formed by the cystines.
And in the end, if we put all that together,
then we come to the realization that the three-dimensional
structure of a protein as determined by the art of x-ray
-- There we go.
I'm not actually dyslexic.
I actually have a cousin who I won't mention
whose son was so dyslexic that when he came to stairways
he didn't know whether to put his foot up or down.
Now that's difficulty.
This is not so bad.
OK, anyhow, because I solved it within less than two minutes
time, all right, so here we see this
is what the three-dimensional structure of a protein
looks like.
This is called a space-filling model
because here, one draws in, as determined
by x-ray crystallography what the,
if we could see what a protein looks
like, what it actually must look like,
where each of the atoms including these side chains
is actually depicted.
Before, when we used these far more schematic descriptions
like here, we were just talking about the overall structure
of the backbone.
We weren't really indicating where the side chains were,
and what space they would fill up.
And, if we give them the chance, if we
put in all of the other atoms, the side chains,
and we create a space-filling model where
the actual atoms are shown, this is
what the protein would look like.
And the fact of the matter is that all virtually proteins
have very specific structures.
It's not as if they can shift from one structure to another.
Once they leave their normal native structure
they will lose their ability to do what their normal jobs are.
And, this particular overhead happens
to bring in yet another theme that we're
going to focus on increasingly, which
is, what do proteins do in cells?
I'm glad I asked that question.
One of the things they do is they act as catalysts, i.e. ,
as enzymes.
The fact is, as we will discuss later,
virtually all biochemical reactions
require an enzyme catalyst in order to propel them forward.
That is to say, if there's a biochemical reaction to occur,
almost always it will not occur spontaneously
the same way that a hydroxyl ion and a hydrogen
will join together spontaneously in water.
Almost all biochemical reactions require the mediation
of an enzyme which is a biological catalyst in order
to encourage this to happen.
And, almost all catalysts in our cells are proteins.
So, if you have 4,326 distinct biochemical reactions occurring
in the cell, that means that there's probably
almost as many distinct enzymes, each one of which
is assigned to mediate one or another
of those distinct biochemical reactions.
And here, we see the fact that this
is an enzyme which happens to be called hexokinase.
Recall that the -ase suffix at the end
dictates that this is already an enzyme rather
than a carbohydrate.
And, this attaches, in fact, a phosphate group onto glucose.
And, what happens is that the glucose, which
is the substrate, which is acted upon by the catalyst,
is pulled into this site in the protein which
is highly specialized to mediate the enzymatic reaction.
Almost all the business of this complex enzyme
is carried out right here.
And somehow, a lot of the other amino acids
that are located at a distance are doing other things
like regulating the activity of the enzyme.
But the actual business end of the enzyme
is present in what is called a catalytic cleft, an active site
of this enzyme in which the substrates are pulled in
and are manipulated and changed chemically
by the actions of this particular enzyme.
Now, in saying that virtually all catalysts, but not all, are
proteins, I also mean to say that proteins
have a second major function in the body.
The first major function is to act as enzymes in catalysts.
The second major function is to create biochemical structures,
i.e., structures of different cytoskeleton proteins such as I
showed you two lectures ago.
And so, we are going to come repeatedly to the situations
where complex structural entities in the cell
are composed of different structural proteins.
Again, this is just a prelude to talking
about these in greater details, these two major functions
of enzymatic catalysis on the one hand,
and creating structure on the other hand.
And so now, we get to really four hierarchical levels
of protein structure.
The primary structure is the amino acid sequence.
And, if we dwell for second on this amino acid sequence,
let's realize that any single amino acid can
follow any other amino acid.
So, what that means is that if glycine
is the first amino acid, as it happens to be here,
serine is only one of 20 different possible
second amino acids.
Aspartic acid is only one of 20 different third amino acids
as the third residue.
We often call these different residues:
the first residue, second residue, third residue,
fourth residue, and fourth, and so forth.
And, keep in mind, if we think about
the combinatorial implications of that,
the first amino acid residue can have 20 different ones.
The second can have 20 different identities.
The third can have 20 different identities.
That means if we make a tripeptide
- a tripeptide has three amino acids in it.
That means we can make 400 dipeptides, 400
distinct dipeptides, and we can make
8,000 distinct tripeptides.
Now, if you imagine that the average amino acid,
the average protein in the cell is,
let's say, 150 amino acid residues long,
that means that in principle, one could make 20
to the 150th power distinct amino acid sequences because
of these absence of any constraints
of which amino acid will follow which other amino acids.
In other words, if the average polypeptide
has this many residues, this is the number
of distinct 150 amino acid residue long proteins
that one could, in principle, synthesize.
I'm not saying all of them have ever
been synthesized since the formation of life
on this planet.
Indeed, since some amino acid chains are 4, 5, 600,
even 2,000 amino acid residues long,
I think the one that is affecting muscular dystrophy
is more than 2,000 amino acids.
Dystrophin.
Does anybody know here?
It's big.
Anyhow, imagine the number of possibilities.
So, combinatorially, life can make almost
whatever types of amino acids it would like by dictating
the sequence of amino acids.
Now, let's just go and look here again.
There is a secondary structure.
The tertiary structure is the way
in which the different alpha helices here
or beta pleated sheets are disposed three-dimensionally
with respect to one another.
And, the quaternary structure represents
how different polypeptides are associated one with the other.
So, for example, hemoglobin is a tetramer.
Hemoglobin doesn't exist as a monomeric protein.
Its solution exists as a tetramer.
And there's two kinds of globin chains.
There is an alpha kind and a beta kind.
And, if we look in a very rough and schematic way at the way
that a hemoglobin tetramer is arranged,
there are two alpha polypeptide chains and two beta polypeptide
chains.
They are not covalently attached to one another.
They are associated with one another via hydrogen bonds
and hydrophobic interactions.
And, this is the actual native configuration
of globin to alpha and to beta chains.
It doesn't exist as a single amino acid in solution.
It exists as a tetramer.
And, indeed, most, or I shouldn't say most,
but very many proteins exist in these configurations
where the tertiary structure represents
four different amino acid chains.
And each of these has an N and C terminal.
Each of these is chemically distinct.
These four could probably be taken apart from one another
simply by raising the temperature.
And, they associate like this.
And, in the absence of this association,
if you just had one of these alphas, or one of these betas,
it wouldn't function well at all.
In fact, it might be totally dysfunctional.
One other thing that may be implicit to you,
but I haven't said, but that is very important to realize
is the following: Let's imagine that this
is the three-dimensional structure of the protein,
as it may well be.
Let's now think about hydrophobic and hydrophilic
amino acids.
The hydrophobic amino acids hate to be present water.
And therefore, they are, we can imagine this case correctly,
tucked away inside the interstices of the protein
far way from the surface.
They don't have any contact with water.
Conversely, the highly charged hydrophilic amino acids
are actually sticking out at the surface.
And this begins to yield yet another insight
into how the three-dimensional stereochemistry of proteins
is maintained and dictated because the hydrophobic amino
acids, through hydrophobic interactions,
stabilize the inner core of the protein that is well
shielded from the aqueous solvent.
The hydrophilic amino acids are on the outside.
They like to be in intimate contact with the water.
So, we already now have talked about a number
of distinct different interactions that
are responsible for creating the three-dimensional
stereochemistry of the protein.
First of all, there are the disulfide bonds,
which create chain-to-chain covalent interactions.
They are the hydrogen bonds in which different chains
can interact one another.
And there are these hydrophobic and hydrophilic interactions.
And, there are some relatively inconsequential van der Waals
interactions, which are really not
worth discussing although some people get
really excited about them.
But we won't.
So, here we now begin to see that we have really interesting
polypeptides unlike the boring polypeptides that
are ultimately the way one must judge carbohydrates.
Some people think carbohydrate chemistry
is really interesting.
But it really isn't that interesting
because you just have the same monomer in a hundred,
or 500 stretches.
Here, a protein is much more interesting
because of this enormous variability
in amino acid sequence, and the consequent ability
to create all kinds of chemical reactivities and structures
because of these 20 different amino acids.
If we were to imagine life on another planet,
and we imagine that there were, let's say,
amino acid-like molecules that were part of life,
maybe that other life wouldn't have exactly the same 20
amino acids as we do.
It almost certainly would be a water base the way we are.
But, it would also rely on hydrogen bonds
and hydrophilicity, and hydrophobicity interactions
in order to dictate the three-dimensional structure.
In the absence of this very specific
three-dimensional structure I will tell you
that this enzyme could not function.
And, if you were to take this enzyme,
if it were typical enzyme and you were to heat it up briefly,
even often slightly above normal body temperature,
it might denature, i.e., it might
lose its three-dimensional structure irreversibly.
And, once it was denatured, this process of denaturation,
it might not be able to spontaneously reassume
that pre-existing three-dimensional configuration
and therefore would forever be inactive.
That means to say very explicitly
that even though the amino acids that
are creating that active catalytic site remain there.
Their highly specific three-dimensional disposition
is critical for the continued actions of this enzyme.
And, once their three-dimensional dispositions
are shifted around through the process of interactions,
then, we have trouble because the enzyme can no longer
do its assigned task.
We are going to go now to an even higher order of complexity
in one sense.
We are going to go to the royalty of the macromolecules,
which are the nucleic acids.
Of course, protein chemists would take great umbrage
at the very notion that there are
things better than proteins.
But, the fact of the matter is, I
can't show you that overhead because it's
from the other textbook which is copyright,
and we are being filmed.
How many people have had the backs
of their heads immortalized on these videos?
Did you call home and ask anybody to identify you?
I don't know, but soon each of us
has the limelight for 15 minutes a lifetime, right?
So, you'll have your 15 minutes.
Here are some nucleic acids.
And let's look at these nucleic acids and the way
they are put together.
Keep in mind to anticipate what we are going to say next time,
once again, we want to make end-to-end aggregates.
We want to polymerize molecules.
And in this case, we want to do so once again
through a dehydration reaction.
And, moreover, just to look at the building blocks
of nucleic acids, we start again in this case with two pentoses.
Recall that they have fuor carbons:
one, two, three, four, did I say four?
You know I meant five.
One, two, three, four, five.
So, whenever I say four from now on,
I mean, or, whenever I say four I may also mean four.
OK, one, two, three, four five.
And let's look at the two basic kinds of pentose molecules
that are present in nucleic acid because they define
the essential difference between DNA and RNA.
Here's a regular old rather familiar kind
of pentose with five carbons.
And here's an unfamiliar kind of pentose
which we call deoxyribose.
Why?
Because if you look really carefully,
you'll see that the hydroxyl group here,
which should be present in any self-respecting pentose is
missing, and is replaced simply by a hydrogen group, i.e.,
it's lost its oxygen, whence cometh the word, ?deoxyribose,?
and ultimately clearly the word, ?deoxyribose nucleic acid.?
And one of the attributes, one of the virtues
of carbohydrates, as we discussed last time,
were these numerous hydroxyl groups,
which represent opportunities for all kinds of dehydration
reactions which can enable one to build
much more complex molecules.
And here, we see the structure of, for example,
a deoxyribonucleotide whose detailed structure
we'll get into next time.
But just, let's look at how these hydroxyl groups
have been used.
The hydroxyl group, in this case in DNA,
and by the way notice that the structure I've shown here,
there's a side chain attached here,
and a side chain attached here.
And neither of those depends on whether or not
there is a hydrogen or a hydroxyl right here.
And look at what's happened here.
Here, we have this hydroxyl over here
to which a base has been attached covalently,
once again, by a dehydration reaction.
And here, we have a situation where actually three phosphate
groups have been attached to the hydroxyl group
in this direction.
And, this represents the basic building blocks
of nucleic acids.
Now, one of the things that's going to be really important
and that you're going to have to memorize, I told you, you
weren't going to have to memorize anything.
But you didn't believe me, did you?
Good.
OK, one of the things you're going to have to memorize
is the numbering system here.
This is number one, two, three, four, five,
or to be totally frank, and you know
I'm always that, one prime, two prime, three prime, four prime,
five prime.
And that numbering system, it turns out,
is going to be very important for our subsequent discussions.
Notice here that, for example, it's here
at the two prime position that this deoxyribose is lacking
the oxygen that is present normally in RNA.
And, with all this in mind, we will wait in great suspense
until Wednesday when we actually talk
about how this is exploited to make highly complex polymers.
Have a good two days.
See you Wednesday at 10:00.