Biochemical methods to map and quantify allosteric motions in human glucokinase.

Autor:	Gordon BH; Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, United States; Institute of Molecular Biophysics, Florida State University, Tallahassee, FL, United States., Liu P; Protein Analytical Chemistry, Genentech Inc., South San Francisco, CA, United States., Whittington AC; Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, United States; Department of Biological Science, Florida State University, Tallahassee, FL, United States., Silvers R; Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, United States; Institute of Molecular Biophysics, Florida State University, Tallahassee, FL, United States., Miller BG; Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, United States. Electronic address: miller@chem.fsu.edu.
Jazyk:	angličtina
Zdroj:	Methods in enzymology [Methods Enzymol] 2023; Vol. 685, pp. 433-459. Date of Electronic Publication: 2023 Apr 19.
DOI:	10.1016/bs.mie.2023.03.009
Abstrakt:	Allosteric regulation of protein function is ubiquitous in biology. Allostery originates from ligand-mediated alterations in polypeptide structure and/or dynamics, which produce a cooperative kinetic or thermodynamic response to changing ligand concentrations. Establishing a mechanistic description of individual allosteric events requires both mapping the relevant changes in protein structure and quantifying the rates of differential conformational dynamics in the absence and presence of effectors. In this chapter, we describe three biochemical approaches to understand the dynamic and structural signatures of protein allostery using the well-established cooperative enzyme glucokinase as a case study. The combined application of pulsed proteolysis, biomolecular nuclear magnetic resonance spectroscopy and hydrogen-deuterium exchange mass spectrometry offers complementary information that can used to establish molecular models for allosteric proteins, especially when differential protein dynamics are involved. (Copyright © 2023 Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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