Probing dimer interface stabilization within a four-helix bundle of the GrpE protein from E. coli via internal deletion mutants: conversion of a dimer to monomer
| Autor: | Andrew F. Mehl, Zohair Ahmed, Aaron Wells, U G Nalin, Tilemahos D. Spyratos |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Models
Molecular Circular dichroism Protein Folding Dimer Biochemistry Article Protein Structure Secondary chemistry.chemical_compound Protein structure Structural Biology Escherichia coli Molecular Biology Protein secondary structure Heat-Shock Proteins DNA Primers Sequence Deletion Helix bundle Protein Stability Circular Dichroism Escherichia coli Proteins General Medicine Crystallography Monomer chemistry Helix Chromatography Gel Protein folding Dimerization Plasmids |
| Popis: | Insight into protein stability and folding remains an important area for protein research, in particular protein-protein interactions and the self-assembly of homodimers. The GrpE protein from Escherichia coli is a homodimer with a four-helix bundle at the dimer interface. Each monomer contributes a helix-loop-helix to the bundle. To probe the interface stabilization requirements, in terms of the amount of buried residues in the bundle necessary for dimer formation, internal deletion mutants (IDMs) were created that sequentially truncate each of the two helices in the helix-loop-helix region. Circular dichroism (CD) spectroscopy showed that all IDM's still contained a significant amount of α-helical secondary structure. IDM's that contained 11 or fewer of 22 residues originally present in the helices, or those that lost at least 50% of residues with less than 20% the solvent accessible surfaces (that is, hydrophobic residues) were unable to form a significant amount of dimer species as shown by chemical cross-linking. Gel filtration studies of IDM3.0 (one that retains 10 residues in each helix) show this variant to be mainly monomeric. |
| Databáze: | OpenAIRE |
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