Hemoglobin v.

Myoglobin
1. Transport and Storage of Oxygen
a. Oxygen required for oxidative metabolism and energy production in
most cells
i. O2 the terminal electron acceptor in aerobic metabolism
ii. Used to oxidize dietary substrates to carbon dioxide, yielding
energy
b. Hemoglobin (Hb) used by vertebrates to transport O2 to tissues from
lung or gills) and to transport CO2 from tissues to lung (or gills)
c. Myoglobin (Mb) used in some tissues, notably muscle, as a storage
reserve of O2 and for intracellular transport of O2
2. Myoglobin vs. Hemoglobin
a. Mb monomeric protein (153 amino acid residues)
b. Hb is a tetramer of 2 alpha chains (141 residues each) and 2 beta
chains (146 residues each)
c. Function of Mb is to bind O2 that has been delivered to tissues by the
hemoglobin in the blood
i. Storing the O2 until its needed as terminal electron acceptor for
energy metabolism, and
ii. Transporting it within the cell (molecular bucket brigade, O 2
dissociates from one Mb molecule and binds to the next)
iii. Function of Mb is to bind O2 with the help of a heme
prosthetic group
iv. Apoprotein + prothetic group holoprotein
v. Heme
1. Prosthetic group made of hydrophobic porphyrin ring
2. Fe2+ is complexed in the porphyrin ring by two main chain
His residues
vi. Oxygen binding changes structure
1. Iron-oxygen bonding
d. Structures are conserved in hemoglobin B and myoglobin
e. Tetramer vs. monomer
i. Hemoglobin is a dimer of dimers
3. Oxygen binding

a.
b. Cooperative binding

i.
4. Binding models
a. Allosteric Cooperativity
b. Concerted Model
i. Only T-state or R-state
c. Sequential Model
i. Both T and R states
ii. KNF
5. Allostery
a. Allosteric regulation change in conformational ensemble upon
binding of an effector, which results in an apparent change in binding
affinity
i. Positive
ii. Negative
1. Non-competitive inhibition
iii. Does NOT require quaternary structure
b. Allosteric regulation of enzymes
c. Oxygen saturation curves for Mb and Hb
i. Storage high binding affinity only
ii. Transport balance between high binding affinity and efficient in
unloading
d. Oxygen binding to Hemoglobin
i. Cooperative binding
ii. Binding of O2 causes conformational change in Hb
iii. Allosteric protein binding of a ligand to one site on the
protein affects the binding properties of another site
e. Other Allosteric Effectors
i. 2,3 BPG
ii. Protons (CO2 or lactic acid)
iii. Homotrophic
1. Substrate is also regulatory
2. Typically an activator
3. Ex: O2
iv. Heterotropic
1. Regulatory molecule that is NOT also the substrate
2. Activator or inhibitor of enzyme
3. Ex: H+, CO2, & BPG are heterotropic modulators of Hb
f. 2,3-Bisphosphoglycerate regulates O2 binding

i. 2,3-BPG another negative heterotropic effector of O2 binding to

Hb
1. Binds strongly to the deoxy form of Hb (T state), but only
very weakly to oxy form (R state)
2. Binds to ONE SITE, in the CENTRAL CAVITY, on the Hb
tetramer
ii. If human hemoglobin did NOT have its O 2 affinity reduced by
bound DPG, we could not effectively release O 2 to our tissues
iii. High Altitude Adaptation
1. Human adaptation to high altitude is a complex
physiological process that involves many events,
including
a. An increase in number of erythrocytes and also
b. An increase in the amount of hemoglobin per
erythrocyte, which may take several weeks to
accomplish
2. One event that occurs within 24 hours is an increase in
the content of 2,3-BPG in the erythrocyte
g. Fetal O2 Transport
i. Fetus obtains O2 by diffusion across placenta
ii. In fetal hemoglobin, the alpha chain is replaced with a gamma
chain
iii. Fetal alpha2gamma2 hemoglobin does not bind 2,3-BPG as well
as adult hemoglobin
h. Sickle Cell Anemia
i. Sickle cell hemoglobin is called hemoglobin S (HbS)
ii. The substituted valine is exposed in deoxyhemoglobin and can
interact with other deoxy HbS to form aggregates that deform
the red blood cells
iii. The sickled cells clog blood flow through the capillaries, leading
to tissue damage
i. Mutant Hemoglobins
i. Provide unique opportunities to probe structure-function
relations in a protein
ii. There are nearly 500 known mutant hemoglobins and >95%
represent single amino acid substitutions
iii. About 5% of the populations carries a variant of hemoglobin
iv. Some mutant hemoglobins casue serious illness
v. The structure of hemoglobin is so delicately balanced that small
changes can render the mutant protein nonfunctional