The Neisseria transcriptional regulator PilA has a GTPase activity

C. G. Arvidson, Magdalene "Maggie" So

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The pilA gene of Neisseria gonorrhoeae encodes the response regulator of a two-component regulatory system that controls pilin gene expression. Examination of the primary sequence of PilA indicates that the protein contains at least two functional domains. The N-terminal region has a proposed helix-turn-helix motif thought to be involved in DNA binding. This region also contains the residues that are presumed to form the acidic pocket involved in phosphorylation by PilB, the sensor kinase of the system. The C terminus of the protein has extensive homology to the G (GTP-binding) domains of the eukaryotic signal recognition particle (SRP) 54-kDa protein and the α subunit of the SRP receptor, or docking protein. This homology also extends to similar regions of the bacterial SRP homologs Ffh and FtsY. Here, we demonstrate that purified PilA has significant GTPase activity, and that this activity has an absolute requirement for MgCl2 and is sensitive to KCl and low pH. We also show that PilA has a strict specificity for GTP, and that GTP hydrolysis follows first order kinetics, with a maximum velocity (V(max)) of 1900 pmol of P(i) produced per min per mg of protein and a K(m) for GTP of 9.6 μM at 37 °C.

Original languageEnglish (US)
Pages (from-to)26045-26048
Number of pages4
JournalJournal of Biological Chemistry
Volume270
Issue number44
DOIs
StatePublished - 1995
Externally publishedYes

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Neisseria
GTP Phosphohydrolases
Guanosine Triphosphate
Signal Recognition Particle
Proteins
Helix-Turn-Helix Motifs
Fimbriae Proteins
Phosphorylation
Magnesium Chloride
Neisseria gonorrhoeae
Protein Subunits
Protein C
Gene expression
Hydrolysis
Phosphotransferases
Genes
Control systems
Gene Expression
Kinetics
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

The Neisseria transcriptional regulator PilA has a GTPase activity. / Arvidson, C. G.; So, Magdalene "Maggie".

In: Journal of Biological Chemistry, Vol. 270, No. 44, 1995, p. 26045-26048.

Research output: Contribution to journalArticle

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AB - The pilA gene of Neisseria gonorrhoeae encodes the response regulator of a two-component regulatory system that controls pilin gene expression. Examination of the primary sequence of PilA indicates that the protein contains at least two functional domains. The N-terminal region has a proposed helix-turn-helix motif thought to be involved in DNA binding. This region also contains the residues that are presumed to form the acidic pocket involved in phosphorylation by PilB, the sensor kinase of the system. The C terminus of the protein has extensive homology to the G (GTP-binding) domains of the eukaryotic signal recognition particle (SRP) 54-kDa protein and the α subunit of the SRP receptor, or docking protein. This homology also extends to similar regions of the bacterial SRP homologs Ffh and FtsY. Here, we demonstrate that purified PilA has significant GTPase activity, and that this activity has an absolute requirement for MgCl2 and is sensitive to KCl and low pH. We also show that PilA has a strict specificity for GTP, and that GTP hydrolysis follows first order kinetics, with a maximum velocity (V(max)) of 1900 pmol of P(i) produced per min per mg of protein and a K(m) for GTP of 9.6 μM at 37 °C.

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