Biochemistry
Biochemistry
8th Edition
ISBN: 9781464126109
Author: Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr., Lubert Stryer
Publisher: W. H. Freeman
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Chapter 7, Problem 14P
Interpretation Introduction

(a)

Interpretation:

An oxygen-binding curve for a hypothetical two subunit hemoglobin with n=1.8 and P50=10 torr should be plotted.

Concept introduction:

Hill equation is represented as follows:

log(Y1Y)=nlog(pO2)nlog(P50)

Here, Y- fractional saturation

n − a measure of the degree of cooperativity in ligand binding.

P50 − partial pressure of oxygen at which hemoglobin is half saturated.

The Hill plot of log(Y/1-Y) versus log(P50) should be a linear graph with slope of n.

Interpretation Introduction

(b)

Interpretation:

An oxygen-binding curve for shypothetical two subunit hemoglobin with n=2, L = 1000, c = 0.01 and KR=1 torr should be plotted using concerted model

Concept introduction:

Concerted model equation is written as follows:

Y=α( 1+α)n1+Lcα( 1+cα)n1( 1+α)n+L( 1+cα)n

Here, Y − fractional saturation

a − ratio between the substrate concentration and the dissociation constant for a ligand binding to a single site in R state.

L − The ratio of the concentrations of the T and R states with no ligands bound

c − the ratio between the dissociation constant for a ligand binding to a single site in R state and that of T state.

n − number of binding sites

Also, the ratio can be calculated as follows:

α=pO2KR

Here,

pO2 − partial pressure of oxygen

KR - the dissociation constant for a ligand binding to a single site in R state.

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In an experiment, hemoglobin is dissociated in a buffer and a subunit is isolated to study for its oxygen binding affinity. (i)  What is the shape of the oxygen dissociation curve is expected in the experiment?Explain why. (ii)  Is the Km of the isolated subunit higher or lower than the Km of an intact hemoglobin?
. Oxygen binding by the hemocyanin of the shrimp Callianassa has been measured. Using the following data, prepare a Hill plot and determine (a) Pso, (b) h (the Hill coefficient), and (c) the minimum number of oxygen binding sites on the protein molecule. Poz (mm Hg) Yoz Po, (mm Hg) Yo2 1.1 0.003 136.7 0.557 7.7 0.019 166.8 0.673 10.7 0.035 203.2 0.734 31.7 0.084 262.2 0.794 71.9 0.190 327.0 0.834 100.5 0.329 452.0 0.875 123.3 0.487 736.7 0.913
(No more than one page). Explain why quaternary structure is necessary for cooperativity and allosteric regulation. Use hemoglobin as an example. Include the various allosteric effectors that influence oxygen binding.
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