The nature of ligand-induced conformational change in transferrin in solution. An investigation using X-ray scattering, XAFS and site-directed mutants

Journal of Molecular Biology
J G GrossmannS S Hasnain

Abstract

Ligand-induced conformational change in transferrins has been studied by site-directed mutagenesis of human serum half molecule (N-lobe), X-ray absorption fine structure (XAFS) spectroscopy and X-ray solution scattering. Use of recent advances in data analysis has been made for extracting model-independent molecular shapes from X-ray solution scattering data for the intact, the half molecule and its mutants. Clear evidence is provided that the transferrin molecule (intact as well as N-lobe), in its apo and holo forms, exists for the majority of the time in well-defined specific conformations representing the "fully opened" and "closed" states of the molecule, respectively. Evidence is also provided for the existence of an additional conformation, referred to here as the "intermediate" conformation for simplicity, which is trapped in the case of some of the mutants in the iron-bound form. We suggest that domain closure in the transferrin molecule is a two-step process, with the intermediate conformation representing the first stage of domain closure (approximately 20 degrees hinge-twist of domain II). Our data are not inconsistent with the ligand-free molecule sampling the closed states occasionally (< or = 10%) but are not in s...Continue Reading

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Citations

Mar 18, 2009·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Shaina L Byrne, Anne B Mason
Oct 28, 2009·The Journal of Biological Chemistry·Jacqueline EllisGordon C K Roberts
Aug 17, 2000·Bioscience, Biotechnology, and Biochemistry·M Hirose
Nov 2, 2004·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·David H HamiltonKenneth N Raymond
Jun 24, 2010·Protein Science : a Publication of the Protein Society·Brian E EckenrothStephen J Everse
Jun 29, 2001·The Journal of Biological Chemistry·K K ChohanM J Sutcliffe
Oct 10, 2002·The Journal of Biological Chemistry·Maolin GuoPeter J Sadler
Jun 5, 2020·Biophysical Journal·David J ReilleyAnastassia N Alexandrova
Dec 26, 2001·Chemical Reviews·H SunP J Sadler
Aug 15, 2009·Chemical Research in Toxicology·Maria G Benavides-Garcia, Krishnan Balasubramanian
Aug 23, 2005·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Lisa A LambertAnne B Mason
Mar 15, 2006·Journal of Immunological Methods·Yasser PereraRolando Pajon
Aug 30, 2011·Biochimica Et Biophysica Acta·Jean-Michel El Hage ChahineNguyêt-Thanh Ha-Duong

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