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PMID: 7961487 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Biochemical characterization of the essential GTP-binding protein Obg of Bacillus subtilis.

Journal of bacteriology ·Vol. 176 ·No. 23 ·1994-12-00 ·Pages 7161-8

Welsh KM, Trach KA, Folger C, Hoch JA

Abstract

An essential guanine nucleotide-binding protein, Obg, of Bacillus subtilis has been characterized with respect to its enzymatic activity for GTP. The protein was seen to hydrolyze GTP with a Km of 5.4 microM and a kcat of 0.0061 min-1 at 37 degrees C. GDP was a competitive inhibitor of this hydrolysis, with an inhibition constant of 1.7 microM at 37 degrees C. The dissociation constant for GDP from the Obg protein was 0.5 microM at 4 degrees C and was estimated to be 1.3 microM at 37 degrees C. Approximately 80% of the purified protein was capable of binding GDP. In addition to hydrolysis of GTP, Obg was seen to autophosphorylate with this substrate. Subsequent release of the covalent phosphate proceeds at too slow a rate to account for the overall rate of GTP hydrolysis, indicating that in vitro hydrolysis does not proceed via the observed phosphoamidate intermediate. It was speculated that the phosphorylated form of the enzyme may represent either a switched-on or a switched-off configuration, either of which may be normally induced by an effector molecule. This enzyme from a temperature-sensitive mutant of Obg did not show significantly altered GTPase activity at the nonpermissive temperature.

Related Genes
obg
MeSH Terms
Bacillus subtilis/enzymology,genetics Bacterial Proteins/genetics,metabolism GTP Phosphohydrolases/genetics,metabolism GTP-Binding Proteins/genetics,metabolism Genes, Bacterial/genetics Genes, Lethal/genetics Guanosine Diphosphate/pharmacology Guanosine Triphosphate/metabolism Hot Temperature Hydrolysis/drug effects Kinetics Models, Biological Molecular Weight Mutation Phosphates/metabolism Phosphoproteins/metabolism Phosphorylation
Chemicals
Bacterial Proteins Obg GTP-binding protein, Bacteria Phosphates Phosphoproteins Guanosine Diphosphate Guanosine Triphosphate GTP Phosphohydrolases GTP-Binding Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Welsh K M
Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037.
Trach K A
Folger C
Hoch J A
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.176.23.7161-7168.1994
Published
1994-12-00
Pages
7161-8
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC197103
Subset
IM
Grants
NIGMS NIH HHS · GM19416 · United States
External Links
PubMed source DOI: 10.1128/jb.176.23.7161-7168.1994 PMC Full Text View journal details
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