PMID: 8955289Dec 1, 1996Paper

Protein conformational change and nucleotide binding involved in regulation of sigmaF in Bacillus subtilis

Journal of Bacteriology
M LordM D Yudkin

Abstract

We have studied the ability of three mutant forms of SpoIIAA, containing amino acid substitutions at the site of phosphorylation (serine 58), to interact with SpoIIAB. Native gel analysis revealed that SpoIIAAS58A could form a complex with SpoIIAB in the presence of ADP and more strongly in the presence of ATP. SpoIIAAS58N did not form a complex with SpoIIAB in the presence of ADP but displayed some interaction with SpoIIAB in the presence of ATP. SpoIIAAS58D was unable to form a complex with SpoIIAB in the presence of either ADP or ATP. Corresponding differences were found in the behavior of the three mutant proteins when studied by gel permeation with high-performance liquid chromatography and limited proteolysis. SpoIIAAS58A behaved like the wild-type SpoIIAA, SpoIIAAS58D like SpoIIAA-P, and SpoIIAAS58N in a way that was intermediate between the behaviors of SpoIIAA and SpoIIAA-P. Limited proteolysis was also used to show that on binding of ADP or ATP SpoIIAB undergoes a shift in conformation. The affinity of SpoIIAB for ADP and ATP was determined by limited proteolysis in the presence of a wide range of nucleotide concentrations. The results indicated that SpoIIAB has approximately equal affinity for ADP and for ATP.

References

Dec 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·R SchmidtR Losick
Feb 15, 1995·Genes & Development·J A Londoño-Vallejo, P Stragier
Mar 14, 1995·Proceedings of the National Academy of Sciences of the United States of America·M L KarowP J Piggot
Mar 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·L Duncan, R Losick
Apr 16, 1996·Proceedings of the National Academy of Sciences of the United States of America·F ArigoniP Stragier
Apr 29, 1996·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·J ErringtonM D Yudkin
Nov 1, 1996·Journal of Bacteriology·S M NajafiM D Yudkin
Jan 1, 1992·Methods in Molecular Biology·D A Harris

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Citations

Sep 3, 2003·Molecular Microbiology·Chien-Cheng ChenOlivier Delumeau
Jan 19, 1999·Annual Review of Microbiology·K T Hughes, K Mathee

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