This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Shiba, T. Articles by Noguchi, T. Search for Related Content PubMed PubMed Citation Articles by Shiba, T. Articles by Noguchi, T. Pubmed/NCBI databases Nucleotide Protein Substance via MeSH

 Previous Article  |  Next Article 

Journal of Bacteriology, March 2005, p. 1859-1865, Vol. 187, No. 5
0021-9193/05/$08.00+0     doi:10.1128/JB.187.5.1859-1865.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Polyphosphate:AMP Phosphotransferase as a Polyphosphate-Dependent Nucleoside Monophosphate Kinase in Acinetobacter johnsonii 210A

Regenetiss Co., Ltd., Okaya,¹ School of Dentistry, Matsumoto Dental University, Shiojiri, Nagano,² Division of Molecular Chemistry, Graduate School of Engineering, Hokkaido University, Sapporo,³ Biochemical Division, YAMASA Corporation, Choshi, Chiba, Japan⁴

Received 1 July 2004/ Accepted 29 November 2004

We have cloned the gene for polyphosphate:AMP phosphotransferase (PAP), the enzyme that catalyzes phosphorylation of AMP to ADP at the expense of polyphosphate [poly(P)] in Acinetobacter johnsonii 210A. A genomic DNA library was constructed in Escherichia coli, and crude lysates of about 6,000 clones were screened for PAP activity. PAP activity was evaluated by measuring ATP produced by the coupled reactions of PAP and purified E. coli poly(P) kinases (PPKs). In this coupled reaction, PAP produces ADP from poly(P) and AMP, and the resulting ADP is converted to ATP by PPK. The isolated pap gene (1,428 bp) encodes a protein of 475 amino acids with a molecular mass of 55.8 kDa. The C-terminal region of PAP is highly homologous with PPK2 homologs isolated from Pseudomonas aeruginosa PAO1. Two putative phosphate-binding motifs (P-loops) were also identified. The purified PAP enzyme had not only strong PAP activity but also poly(P)-dependent nucleoside monophosphate kinase activity, by which it converted ribonucleoside monophosphates and deoxyribonucleoside monophosphates to ribonucleoside diphosphates and deoxyribonucleoside diphosphates, respectively. The activity for AMP was about 10 times greater than that for GMP and 770 and about 1,100 times greater than that for UMP and CMP.

This article has been cited by other articles:

• Zhang, H., Rao, N. N., Shiba, T., Kornberg, A. (2005). Inorganic polyphosphate in the social life of Myxococcus xanthus: Motility, development, and predation. Proc. Natl. Acad. Sci. USA 102: 13416-13420 [Abstract] [Full Text]