Welcome to Scribd, the world's digital library. Read, publish, and share books and documents. See more
Standard view
Full view
of .
Look up keyword
Like this
0 of .
Results for:
No results containing your search query
P. 1
Week Four Lecture 560B on Line

Week Four Lecture 560B on Line

Ratings: (0)|Views: 33|Likes:
Published by TheNourishedSprout

More info:

Published by: TheNourishedSprout on Feb 07, 2012
Copyright:Attribution Non-commercial


Read on Scribd mobile: iPhone, iPad and Android.
download as RTF, PDF, TXT or read online from Scribd
See more
See less





Week Four LectureBiochemistry of NutritionDr. Charles SaladinoIntroductory Concepts
We could begin this lecture by immediately describing the structure of various proteins.However, in order to understand the relationship between the structure and the functionof proteins, such as hemoglobin (Hb), it is necessary that we comprehend the criticalconcept of allosterism, derived from the Greek 
(other) +
(solid or space).Although this term is a key feature of many enzymes, it is certainly not limited to these biomolecules, but rather, is critical to the function of many other types of proteins.Before we start with the classic model of hemoglobin to illustrate allosterism, we need todefine the term as involving the behavior of multisubunit (thus having quaternarystructure) proteins that bind small molecules. This binding of a small molecule, called aligand, to the protein will change the affinity of that protein for other ligand moleculesand/or the activity level of the protein itself. In other words, allosteric effects involve the binding of a ligand at one site, which in turn affects the binding of a ligand at another site(or subunit) in the same protein, probably altering the protein’s activity. To accomplishthis, there must be conformational shifts involving interactions between the subunits of the protein. (The conformational shifts involved in some proteins are not wellunderstood, whereas those of others, including hemoglobin, are very well described, aswe shall see.)The following are the main features of the symmetry model of allosterism, firstformulated in 1965. However, I have included the more recent modifications to thisoriginal model:1. The protein must be an oligomer (meaning having multiple subunits) of symmetrically-related subunits (or approximately symmetrically related - as wewill discuss in
the case of the α- and β-subunits of Hb).
2. The oligomer (multi-subunit protein) can exist in two conformational states,Taught (T) and Relaxed (R), which exist in equilibrium with each other.3. A ligand binds to a subunit that is configured to accept it. This binding resultsin a change in the conformation of that subunit, which, in turn, initiates a changein the conformation of a second subunit to alter that second subunit’s affinity for the ligand (and the activity level of the protein– to be discussed under the topic of enzymes).4. In other words, when the first ligand binds to the first subunit, the rest of thesubunits of the protein change conformation in a sequential manner (notsimultaneously as previously thought), so as to accept additional ligandssequentially and generally change the activity of the whole protein.5. As a result of the sequential change in the conformation of each subunit, theentire protein will go from the Taught State to the Relaxed State or vise versa,depending upon whether you are starting with the protein in the R State or the T
State of configuration. Classically, the T form of the protein is less active,whereas the R form is more active.As we shall see, this phenomenon of allosterism allows for efficient and multiple ways of regulating the ability of a protein to bind a ligand and to become either more active (R form) or less active (T form). Those ligands that bind to the protein or to an enzyme toenhance its function are known as positive allosteric effectors, whereas those that bind tocause a decreased level of functional are referred to as negative allosteric effectors.
Structure of Hemoglobin (Hb)
When the metabolic pathways associated with aerobic respiration began to appear, so didthe ability of cells to produce great sums of additional energy, compared to the relativelysmall amount of ATP that could be generated from anaerobic respiration. With higher levels of energy production, greater and greater molecular-genetic variability could besupported, and a huge leap in specialized cellular activity could be realized. However, asystem had to be available to deliver that insoluble oxygen to the tissues for aerobicrespiration. The molecular and cellular environment within the erythrocyte provided justwhat was needed for delivering oxygen from the lungs to the tissues, within the contextof the circulatory system. Within this intra-erythrocytic environment, wherein there areneither nuclei nor mitochondria, are found the well described hemoglobin molecules ingreat quantity plus an effective bicarbonate buffer system, which we shall discuss shortlyin detail. The erythrocyte is the only cell that contains hemoglobin.The primary function of hemoglobin is to transfer oxygen from the lungs to the tissues. Itis a tetrameric, heme protein that is comprised of four subunits (hence tetrameric), twoidentical
subunits and two identical
β-subunits. Thus,
Hb has quarternarystructure (remember, quarternary refers to a protein having two or more subunits). Boththe
- and the
-subunits are structurally related to one another and to the single subunitmolecule, myoglobin (which we discuss later in this lecture), even though only about18% of the amino acids are identical in the twp types of subunits.
Each α-subunit iscomprised of 141 amino acids, whereas each β-subunit is composed of 146. About 75% of the complete structure of hemoglobin comprised of these four subunits is in the form of the α-helix.
Within the center of eachof the four subunits is a heme molecule, which is a protoporphyrin ring structure with aniron atom in the center.
Familiarize yourself with the heme structure, shown on page355 of your text, for the second exam.
Below is a ribbon diagram of the hemoglobinmolecule showing the four subunits, each in a different color and with each containing aheme molecule. As you observe the diagram,
consider why each subunit fulfills thecriteria of having tertiary structure (review those criteria from last week)
and howthe bonding of those subunits forms a quaternary protein structure.
The iron atom at the center of the heme is coordinately bonded with four nitrogen atomsand is the actual binding site of oxygen to the hemoglobin molecule
see page 355 of yourtext)
. Thus, one hemoglobin molecule binds four oxygen molecules. However, the issueis exactly when and how the oxygen molecules bind to the hemoglobin in the lungs andwhen and how they are released to the tissues. Obviously, the oxygen gradients that existin the lungs vs. the tissues allow the diffusion of oxygen between the red blood cells andthe tissues and the lungs and these erythrocytes. However, the mechanism by whichoxygen binds to and is released from a Hb molecule involves the complexities of allosterism and a specific aspect of it called positive cooperativity. Let’s explain that. Now try and picture one alpha subunit and one beta subunit joining to form a dimer withthe help of numerous hydrophobic interactions.
(This is shown in Devlin. on page 361)
However, Hb is obviously composed of two dimers. One dimer (an α-and a β-subunit) will then interact with the other dimer (the other α-and β-subunit)
, however, by a small number of relatively weak bonding interactions.This is so that one dimer can rotate about 15 degrees in relation to the other dimer whenoxygen binds to the first subunit. So let’s put all this together and define the allostericchange.We begin with the Hb in the Taut state configuration. The first oxygen binds to ahemoglobin molecule while the red cell is in the lungs. This causes an alloseteric,conformational change in the second subunit, which now accepts the second oxygen.This, in turn, repeats as it sequences to the third and then to the fourth subunit, duringwhich time the 15 degree rotation has occurred between the two dimers. The Hb is nowin the relaxed state (R), and it is fully saturated with four oxygen molecules, each boundto an Fe atom in the center of each heme. Obviously, there have been allosteric changeswithin each Hb as oxygen molecules bind. This change in the conformation of onesubunit enhancing the binding of oxygen to the next subunit (and so forth) is defined as“positive cooperativity.” The red cells now travel to the tissues where oxygen is released. When the red cells reach the tissues, the pH is somewhat lower (more protons present),due to metabolic activity and due to carbonic acid (see below). These protons displacethe oxygen from the Hb by
their binding to the Hb in such a manner as tochange the position of the heme relative to the α or β subunit
. In other words, the protons cause displacement of the oxygen molecules as those protons bond tothe heme. This removal of oxygen from the Hb by protons is known as the Bohr Effect.You might have thought that carbon dioxide would take the place of oxygen as it diffusesinto the erythrocyte. Actually, only about 20% of the CO
is carried back to the lungs

You're Reading a Free Preview

/*********** DO NOT ALTER ANYTHING BELOW THIS LINE ! ************/ var s_code=s.t();if(s_code)document.write(s_code)//-->