Hemoglobin: Portrait of a Protein in Action

7.3 H+ ions and CO2 Promote the Release of Oxygen: The Bohr Effect

Rapidly metabolizing tissues general large amounts of H+ and CO2 . To release O2 where the need is greatest, hemoglobin has evolved to respond to higher levels of these substance. H+ and CO2 are allosteric effectors of hemoglobin The regulation of O2 binding by H+ and CO2 is call Bohr effect (1904)

The oxygen affinity of Hemoglobin as pH (7.4 7.2): tendency to release oxygen increases For example: Transport The lungs (pH 7.4, 100 torr) Active muscle (pH 7.2, 20 torr): Result: in a release of oxygen 66% (pH 7.4) 77% (pH 7.2)

At least two sets of chemical groups are important for sensing changes in pH: -amino groups of a chain N-term side chains (His) of b146 and a122 pKa values near 7

 In deoxyhemoglobin, the terminal carboxylate group of 146 forms a salt bridge with lysine residue in the subunit of the other dimer. This interaction locks the side chain of histidine 146 in a position from which it can participate in a salt bridge with negatively charged aspartate 94 in the same chain The formation of these salt bridges stabilizes the T state leading to a greater tendency for oxygen to be release.
H-bond between protonated His 146 and 94 Asp stabilize the salt bridge

In deoxyhemoglobin, three amino acids form two salt bridges that stabilize the T state quaternary structure. Green dash line: hydrongen bond

CO2 stimulates oxygen release by two mechanisms First, high concentrate of CO2 leads to a drop in pH within red blood cell (RBC) Second, direct chemical interaction between CO2 and Hb stimulates oxygen release
carbonic anhydrase

H2CO3: pKa 3.5

CO2 decrease the affinity of Hb for oxygen

CO2 stabilizes deoxyHb by reacting with the terminal amino group to form carbamate group (- charged) terminal amino group lie at the interface between the ab dimers salt bridge stabilizes the T state (favoring the release of O2) a mechanism of CO2 transport (~14%)

 CO2 released from RBC lung (in the form of HCO3-) exchange of HCO3- and Cl- through antiporter

Figure: transport of CO2 from tissues to lungs. Most carbon dioxide is transported to the lungs in the form of HCO3- produced in red blood cells and then release into the plasma. A lesser amount is transport bay hemoglobin in the form of an attached carbamate

7.4 Mutations in Gene Encoding Hb Subunits Can Result in Disease

The notion that: disease might be caused by molecular defects Linus Pauling (1949) to explain the blood disease sickle cell anemia. The name of the disorder the abnormal sickle shape of red blood cells deprived of oxygen

 Sickle-cell anemia results form the aggregation of mutated deoxyHb molecules: Hb molecules have formed large fibrous aggregates

Figure: Sickle-cell hemoglobin fibers

 Examination of the structure of hemoglobin S reveals that the new valine residue lies on the surface of the T-state molecule.

decreased the solubility of deoxy Hb : Val6-Phe85, Lue88 aggregation HbS fiber is formed from 14 chains of multiple interlinked Hbs

Why do these aggregates not form when HbS is oxygenated? : oxyHb in R state Phe 85, Leu88 on b chain are largely buried inside the Hb assembly

Figure: Sickle-cell trait and malaria A significant correlation is observed between regions with a high frequency of the HbS allele and regions with a high prevalence of malaria

Thalassemia is caused by an imbalanced production of hemoglobin chains Thalassemia, the order prevalent inherited disorder of hemoglobin , is caused by the loss or substantial reduction of a single hemoglobin chain. Thelassemia is a set of related diseases, - In Thelassemia the chain of hemoglobin is not produced in sufficient quantity - In Thelassemia the chain of hemoglobin is not produced in sufficient quantity

The accumulation of free alpha-hemoglobin chain is prevented AHSP: -hemoglobin stabilizing protein, in RBCs The ASHP binds to the chain monomers and creates a highly soluble complex; does not precipitate AHSP: - It can bind to both the oxygenated and deoxygenated forms of the alpha chain - Binds to the alpha chain as it is being produced -It is displaced when beta-hemoglobin is produced -Prevents accumulation of free alpha hemoglobin

Additional globin are encoded in the human genome In additional to the gene for myoglobin, the two gens for -hemoglobin, and the one for hemoglobin, the human haploid genome contain other globin genes. Examination of the human genome sequence has revealed two additional globins: neuroglobin, cytoglobin

Thank you for your listening

Question
1. The amount of oxygen released by the hemoglobin molecules in the blood to the tissues a. decreases as temperature increases b. increases as blood pH decreases(acidity increases) c. decreases as pCO2 increases d. increases as pCO2 decreases B

2. Most carbon dioxide is transported as __________ in the blood. bicarbonate ions

3. Which of these factors increases respiratory rate (increases release O2 )? A)increased blood pCO2 B)increased blood pH C)increased blood pO2 D)all of these A

4. What do the three amino acids in deoxyhemoglobin form in order to stabilize the T state quaternary structure? salt bridges

5. How does carbon dioxide form carbamate?

by reacting with the terminal amino group

6. What is the reason thalassemia occurs? by an imbalanced production of hemoglobin chains