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.
|Authors||Haward, S. J., Shewry, P. R., Marsh, J., Miles, M. J. and Mcmaster, T. J.|
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.
|Keywords||Anatomy & Morphology; Biology; Microscopy|
|Year of Publication||2011|
|Journal||Microscopy Research And Technique|
|Journal citation||74 (2), pp. 170-176|
|Digital Object Identifier (DOI)||doi:10.1002/jemt.20888|
|Open access||Published as non-open access|
|Funder||Biotechnology and Biological Sciences Research Council|
|Funder project or code||Centre for Crop Genetic Improvement (CGI)|
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