Physiol-10B15 Discuss the role of haemoglobin as a buffer.
Background
Haemoglobin = iron-containing metalloprotein abundant in red blood cells,
responsible for oxygen transport
Buffer = a substance (usu. weak acid or base) which resists large pH changes from
the addition of another acid or base
e.g. H+ + B → HB+
H+ is absorbed by buffer B to form HB+, which will minimally affect pH
Factors required for effective buffering
Efficiency of buffer increased with:
(1) higher [buffering agent]
(2) pKa of buffering agent matches pH of buffer solution
(3) buffering system is open (e.g. HCO3–/CO2 system is open as the dissolved
[CO2] is able to be varied rapidly via ventilation)
Hb as a buffer
(1) Hb is present in large quantities within RBC (~ 140 g/L, which is 2×[plasma
protein])
(2) Hb contains large number (38) of histidine residues with pKa ≈ 6.8, which
closely match plasma pH ! act as effective buffer
(3) When Hb is deoxygenated ! conformational change ! histidine group pKa
increases from 6.8 to 7.9 ! acid buffering capacity improves
(4) Hb can also bind CO2 ! form carbamino compound ! buffers + carries CO2
Clinical importance:
- buffers H+ and CO2 formed from cellular metabolism
- buffers H+ load from fixed acids ingestion, prior to its renal excretion
- assists CO2 carriage, especially in venous blood
Reactions within RBC
(1) cellular metabolism produces CO2
(2) CO2 produced is taken up by RBC
(3) CO2 is carried by Hb ! carbamino-Hb compound ! produces H+ ! buffered
by Hb histadine groups (this accounts for 30% of venous CO2 carriage)
(4) H2O + CO2 → carbonic anhydrase → H2CO3 ! H+ + HCO3–
(5) Hb acts as a buffer for H+ (deoxyHb better than oxyHb) ! this accounts for
30% of Haldane effect
(6) HCO3– in RBC exchanged for Cl– via antiporter ! Hamburger effect
Examiner’s Comments – pass rate was 27%.
Marks were awarded for a buffer definition, a brief description of haemoglobin
structure, and listing the factors that create an effective buffer. Haemoglobin acts
as an effective buffer for CO2 due to the solubility of CO2, presence of carbonic
anhydrase, and the capacity of haemoglobin.
Important points that needed to be covered included; the importance of the large
volume of haemoglobin, the role and amount of the imidazole side chains on the
histidine residues, the effect that their pKa (6.8) has on buffering ability, and a clear
description of the reactions within the red cell that involves CO2 conversion to
HCO3- and H+ and the fate of these two products.
Discussion about why the deoxy form of haemoglobin is a better buffer due to it’s
change in pKa (7.9), and that this occurs where buffering is needed most, which limits
the pH drop for venous blood. Some candidates correctly identified that the
dissociation of carbamino compounds within the red cell adds H+ which then
needs to be buffered.
Discussing the haemoglobin oxygen dissociation curve, carbon dioxide dissociation
curve, and other buffers in the body did not attract marks.