Identification of a novel nucleoside triphosphatase from Mycoplasma mobile: a prime candidate motor for gliding motility.
A protein with a molecular mass of 42 kDa (P42) from Mycoplasma mobile, one of several mycoplasmas that exhibit gliding motility, was shown to be a novel NTPase (nucleoside triphosphatase). Although the P42 protein lacks a common ATP-binding sequence motif (Walker A), the recombinant proteins expressed in Escherichia coli certainly hydrolysed some nucleoside triphosphates, including ATP. The results of photoaffinity labelling by an ATP analogue supported that the P42 protein contains a specific binding site for ATP (or another nucleoside triphosphate). In the M. mobile genome, the P42 gene is located downstream of gli123, gli349 and gli521 genes, and they have been reported to be polycis-tronically transcribed. As the huge proteins encoded by gli123, gli349 and gli521 play a role in gliding motility of M. mobile, P42 might also have some kind of function in the gliding motility. The gliding motility of M. mobile is driven directly by ATP hydrolysis, but the key ATPase has not been identified. Our results showed that, among these four proteins, only P42 exhibited ATPase activity. Biochemical characteristics--optimal conditions for activity, substrate specificities, and inhibiting effects by ATP analogues--of the recombinant P42 proteins were very similar to those of a putative ATPase speculated from a previous analysis with a gliding 'ghost' whose cell membrane was permeabilized by Triton X-100. These results support the hypothesis that the P42 protein is the key ATPase in the gliding motility of M. mobile.
Ohtani
Naoto
N
Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka 558-8585, Japan. ohtani@ttck.keio.ac.jp
Miyata
Makoto
M
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology*; D003001:Cloning, Molecular; D017931:DNA Primers; D007700:Kinetics; D009174:Mycoplasma / Q000201:enzymology* / Q000502:physiology*; D043583:Nucleoside-Triphosphatase / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity
D001426:Bacterial Proteins; D017931:DNA Primers; D011994:Recombinant Proteins; D043583:Nucleoside-Triphosphatase