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PMID: 12426333 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Aquifex aeolicus PilT, homologue of a surface motility protein, is a thermostable oligomeric NTPase.

Journal of bacteriology ·Vol. 184 ·No. 23 ·2002-12-00 ·Pages 6465-71

Herdendorf TJ, McCaslin DR, Forest KT

Abstract

Bacterial surface motility works by retraction of surface-attached type IV pili. This retraction requires the PilT protein, a member of a large family of putative NTPases from type II and IV secretion systems. In this study, the PilT homologue from the thermophilic eubacterium Aquifex aeolicus was cloned, overexpressed, and purified. A. aeolicus PilT was shown to be a thermostable ATPase with a specific activity of 15.7 nmol of ATP hydrolyzed/min/mg of protein. This activity was abolished when a conserved lysine in the nucleotide-binding motif was altered. The substrate specificity was low; UTP, CTP, ATP, GTP, dATP, and dGTP served as substrates, UTP having the highest activity of these in vitro. Based on sedimentation equilibrium and size exclusion chromatography, PilT was identified as a approximately equal 5- to 6-subunit oligomer. Potential implications of the NTPase activity of PilT in pilus retraction are discussed.

MeSH Terms
Acid Anhydride Hydrolases/chemistry,genetics,isolation & purification,metabolism Adenosine Triphosphatases Amino Acid Sequence Bacteria/enzymology,genetics Bacterial Proteins/chemistry,genetics,isolation & purification,metabolism Cloning, Molecular Dimerization Enzyme Stability Hot Temperature Molecular Motor Proteins Molecular Sequence Data Nucleoside-Triphosphatase Sequence Homology, Amino Acid Substrate Specificity
Chemicals
Bacterial Proteins Molecular Motor Proteins Acid Anhydride Hydrolases Adenosine Triphosphatases Nucleoside-Triphosphatase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Herdendorf Timothy J
Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
McCaslin Darrell R
Forest Katrina T
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.184.23.6465-6471.2002
Published
2002-12-00
Pages
6465-71
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC135430
Subset
IM
Grants
NIGMS NIH HHS · R01 GM059721 · United States
NIGMS NIH HHS · GM59721 · United States
External Links
PubMed source DOI: 10.1128/JB.184.23.6465-6471.2002 PMC Full Text View journal details
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