Analysis of the order of free energy couplings between ligand binding and subunit assembly in human hemoglobin
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Updated: Aug 13, 2026

T-wave Ion Mobility-mass Spectrometry: Basic Experimental Procedures for Protein Complex Analysis
Published on: July 31, 2010
Abstract:
The concept of free energy couplings has been extensively used in studies of the ligand-linked subunit assembly of oligomeric proteins such as human hemoglobin A [cf. Ackers, G. K. (1980) Biophys. J. 32, 331-346]. Recently, the concept of "order" of free energy couplings has been introduced as a description of the number of protein subunits that must be liganded to effect changes in intersubunit interactions [Weber, G. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 7098-7102]. That report utilized the concept of order of free energy couplings to analyze a set of previously published equilibrium constants derived from data pertaining to the chemical equilibrium between oxygen and stripped hemoglobin A [Mills, F. C., Johnson, M. L., & Ackers, G. K. (1976) Biochemistry 15, 5350-5362]. The Weber report claims to have "unequivocally" demonstrated that the coupling between oxygenation and subunit assembly in hemoglobin A is "first order". In the present report, it is demonstrated that free energy couplings of both the first and second order are capable of describing the original oxygen binding data.
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