Transfer of human alpha- to beta-hemoglobin via its chaperone protein: evidence for a new state

The Journal of Biological Chemistry
Véronique Baudin-CreuzaMichael C Marden

Abstract

The alpha-hemoglobin-stabilizing protein (AHSP), a small protein of 102 amino acids, is synthesized in red blood cell precursors. It binds specifically to alpha-hemoglobin (alpha-Hb) subunits acting as a chaperone protein, preventing the formation of alpha-hemoglobin-cytotoxic precipitates. We have engineered recombinant AHSP in a pGEX vector to study the functional consequence of interaction between AHSP and alpha-Hb. By in vitro binding assays, we have isolated the complexes glutathione S-transferase-AHSP.alpha-Hb and AHSP.alpha-Hb. The latter assembles as a heterodimer based on size-exclusion chromatography. These complexes exhibited monophasic CO binding kinetics, as observed for isolated alpha- and beta-subunits of hemoglobin. However, the rate of CO (or oxygen) binding to alpha-hemoglobin bound to its chaperone is three times slower than that observed for isolated alpha-hemoglobin, demonstrating a form that is intermediate to the R- and T-hemoglobin states. The physiologically relevant replacement of the chaperone by beta-hemoglobin chains could be detected by both ligand binding kinetics and tryptophan fluorescence quenching.

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Citations

Sep 19, 2006·Journal of the American Society for Mass Spectrometry·Brian L Boys, Lars Konermann
Apr 8, 2010·The Journal of Biological Chemistry·Thomas BrilletMichael C Marden
Jan 31, 2012·The Journal of Biological Chemistry·Eugene KhandrosMitchell J Weiss
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Jun 2, 2009·Clinical Biochemistry·Corinne VasseurVéronique Baudin-Creuza
Dec 13, 2005·Annals of the New York Academy of Sciences·Mitchell J WeissAndrew J Gow
Sep 26, 2013·Biochimica Et Biophysica Acta·Laurent KigerVéronique Baudin-Creuza
Mar 1, 2015·Transfusion clinique et biologique : journal de la Société française de transfusion sanguine·C Vasseur, V Baudin-Creuza
Jan 5, 2006·Protein Engineering, Design & Selection : PEDS·Corinne Vasseur-GodbillonVéronique Baudin-Creuza
Jun 24, 2017·British Journal of Haematology·Corinne VasseurVéronique Baudin-Creuza
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Aug 12, 2006·The Journal of Biological Chemistry·Suiping ZhouAndrew J Gow

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