HAEMOGLOBIN
Hemoglobin
Koshland Nemethy Filmer model
Crystal structure given by Max Perutz
HbA= 2 α+2β subunits ; & identical 3-D structure.
Bound prosthetic group called heme; red color of blood.
Organic component and a central iron atom.
Organic component : protoporphyrin ; four pyrrole rings linked by methene
bridges to form a tetrapyrrole ring.
Four methyl groups, two vinyl groups, and two propionate side chains are
attached.
Cooperativity Enhances Oxygen Delivery by Hemoglobin.
Binding sites, hemoglobin delivers more O2 to tissues than would a noncooperative
protein
Binding of the oxygen molecule at the sixth coordination site of the iron ion substantially
rearranges the electrons within the iron so that the ion becomes effectively smaller
Change in electronic structure ---- magnetic properties of hemoglobin functional
magnetic resonance imaging
Oxygen Binding Markedly Changes the Quaternary Structure of Hemoglobin
Pair of identical a b dimers (α1β1 and α2β2)
Deoxyhemoglobin, these a b dimers are linked by an extensive interface, which
includes, among other regions, the carboxyl terminus of each chain.
Heme groups separated in the tetramer with iron-iron distances ranging from 24 to
40 Å.
Transition from T to R State in Hemoglobin
Deoxy form = T form
On oxygen binding, T-to-R state transition
Interface between dimers is most effected by this structural transition.
Histidine residue bound in the fifth coordination site
Oxygen binds to hemaglobin
structural transition
iron ion moves into the plane of the porphyrin
carboxyl terminal end of this histidine bound αhelix lies in the interface between the two αβ dimers
transmission to the other subunits
Oxygen Affinity of Hemoglobin
Oxygen affinity of purified hemoglobin > hemoglobin within red blood
cells
Highly anionic compound is present in red blood cells= haemoglobin
Without 2,3-BPG, releasing only 8% of its cargo in the tissues.
2,3-BPG affect oxygen affinity- HOW?
Crystal structure reveals 2,3-BPG binds in a pocket, present only in the T form
T-to-R transition
pocket collapses
2,3-BPG binding to hemoglobin has crucial physiological consequences
fetal hemoglobin two α chains and two γ chains
Substitution of a serine residue for His 143 in the b chain of the 2,3-BPG-binding site
Removes two positive charges from the 2,3-BPG-binding site
Mode of Binding of 2,3-BPG to Human Deoxyhemoglobin
oxygen-binding affinity of fetal hemoglobin > maternal (adult) hemoglobin
Carbon dioxide also stabilizes deoxyhemoglobin by reacting with the terminal amino
groups to form carbamate groups, which are negatively charged.
The amino termini lie at the interface between the a b dimers, and these negatively charged
carbamate groups participate in saltbridge interactions, characteristic of the T-state
structure, which stabilize deoxyhemoglobin's structure and favor the release of oxygen
The Bohr Effect: Hydrogen Ions and Carbon Dioxide Promote the Release of Oxygen
oxygen affinity of hemoglobin decreases as pH (7.4) decreases
Lungs pH 7.4 & pO2 100 torr pH 7.2 pO2 20 torr
Hb
release of oxygen amounting to 77%
The regulation of oxygen binding by hydrogen ions and carbon dioxide is called the Bohr effect
2 sets of chemical groups are responsible for the effect of protons: the amino termini and the side chains of histidines
β 146 and α122, which have pK a values near pH 7.
Consider histidine β146.
In deoxyhemoglobin, the terminal carboxylate group of β146 forms a salt bridge with a lysine residue in the α
subunit of the other dimer.
Side chain of histidine β 146 : salt bridge with negatively charged aspartate 94 in the same chain, provided that the
imidazole group of the histidine residue is protonated
At high pH, the side chain of histidine b146 is not protonated and the salt bridge does not form.
As the pH drops, however, the side chain of histidine b146 becomes protonated, the salt bridge
with aspartate b94 forms, and the quaternary structure characteristic of deoxyhemoglobin is stabilized, leading to a
greater tendency for oxygen to be released at actively metabolizing tissues.