The cytoplasmic fragment of the aspartate receptor displays globally dynamic behavior

Biochemistry
S K SeeleyL K Thompson

Abstract

A number of cloned soluble fragments if the bacterial chemotaxis transmembrane receptors retain partial function. Prior studies of a fragment corresponding to the cytoplasmic domain (c-fragment) of the Escherichia coli aspartate receptor have correlated the signaling state of mutant receptors with the oligomerization state of the c-fragments: equilibria of smooth-swimming mutants are shifted toward oligomeric states; tumble mutants are shifted toward monomeric states [Long, D. G., & Weis, R. M. (1992) Biochemistry 31, 9904-9911]. We have applied several experimental probes of local and global structural flexibility to two signaling states, the wild-type (monomeric) and S461L smooth mutant (predominantly dimeric) c-fragments. Featureless near-UV CD spectra are observed, which indicate that the single Trp residue is in a symmetric environment (most likely averaged by fluctuations) and suggest that the C-termini of both proteins are highly mobile. Both proteins undergo extremely rapid proteolysis and enhance ANS fluorescence, which indicates that many sites are accessible to trypsin cleavage and hydrophobic sites are accessible to ANS binding. The global nature of the flexibility is demonstrated by 1H NMR studies. Lack of chemical...Continue Reading

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Citations

May 31, 2001·Journal of Molecular Graphics & Modelling·A K DunkerZ Obradovic
Sep 26, 1997·Genes & Development·A S CarrollT K Blackwell
May 22, 2002·Protein Science : a Publication of the Protein Society·Alexander BarnakovGerald L Hazelbauer
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Oct 13, 2006·Proceedings of the National Academy of Sciences of the United States of America·Ivan L BudyakJudith Klein-Seetharaman
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Jan 1, 2008·Trends in Biochemical Sciences·Gerald L HazelbauerJohn S Parkinson
Aug 11, 2015·Protein Science : a Publication of the Protein Society·Nicholas L Bartelli, Gerald L Hazelbauer
Jan 21, 2004·Molecular Microbiology·Michael W Bunn, George W Ordal
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Mar 2, 1999·Journal of Structural Biology·S L Mowbray, M O Sandgren
Sep 13, 2001·The Journal of Biological Chemistry·O J MurphyL K Thompson

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