Force spectroscopy of an elastic peptide: effect of D2O and temperature on persistence length

A - Papers appearing in refereed journals

Haward, S. J., Shewry, P. R., Marsh, J., Miles, M. J. and Mcmaster, T. J. 2011. Force spectroscopy of an elastic peptide: effect of D2O and temperature on persistence length. Microscopy Research And Technique. 74 (2), pp. 170-176.

AuthorsHaward, S. J., Shewry, P. R., Marsh, J., Miles, M. J. and Mcmaster, T. J.
Abstract

This study explores the mechanical unfolding of elastic protein analogues as a function of temperature, in both H2O and D2O, using atomic force microscopy (AFM) force spectroscopy in a specially constructed AFM liquid cell. This represents the first time that the effect of D2O on protein flexibility has been investigated at the single molecule level by this technique. Model elastic peptides, R6, were encoded from synthetic genes expressed in Escherichia coli. The peptides possess short N- and C-terminal domains with central repetitive domains containing 13 repeats of the motif PGQGQQ-plus-GYYPTSLQQ. These sequences mimic those in native high molecular weight subunit glutenin proteins which confer elasticity to bread dough. Fitting single molecule stretching events to the worm-like chain model, allows determination of the molecular persistence length under various experimental conditions. The effect of increasing the temperature is to increase the persistence length of the peptide in both H2O and D2O, consistent with the expected "thermal softening" effect. However, the effect is significantly enhanced in D2O, in which the persistence length at 45 degrees C is similar to 25% greater than the value measured in H2O at the same temperature. Stronger intrapeptide H-bonding due to isotopic substitution of hydrogen for deuterium is the most likely cause of the enhanced backbone rigidity. Microsc. Res. Tech. 74:170-176, 2011. (C) 2010 Wiley-Liss, Inc.

KeywordsAnatomy & Morphology; Biology; Microscopy
Year of Publication2011
JournalMicroscopy Research And Technique
Journal citation74 (2), pp. 170-176
Digital Object Identifier (DOI)doi:10.1002/jemt.20888
PubMed ID21275005
Open accessPublished as non-open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeCentre for Crop Genetic Improvement (CGI)
ISSN1059910X
1059-910X
PublisherWiley

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