1887

Abstract

Putrescine can be synthesized either directly from ornithine by ornithine decarboxylase (ODC; the product) or indirectly from arginine via arginine decarboxylase (ADC; the product). The authors identified the and genes in PAO1. The activities of the two decarboxylases were similar and each enzyme alone appeared to direct sufficient formation of the polyamine for normal growth. A mutant defective in both and was a putrescine auxotroph. In this strain, agmatine deiminase (the product) and -carbamoylputrescine amidohydrolase (the product), which were initially identified as the catabolic enzymes of agmatine, biosynthetically convert agmatine to putrescine in the ADC pathway: a double mutant of and was a putrescine auxotroph. AguA was purified as a homodimer of 43 kDa subunits and AguB as a homohexamer of 33 kDa subunits. AguA specifically deiminated agmatine with and values of 0·6 mM and 4·2 s, respectively. AguB was specific to -carbamoylputrescine and the and values of the enzyme for the substrate were 0·5 mM and 3·3 s, respectively. Whereas AguA has no structural relationship to any known C–N hydrolases, AguB is a protein of the nitrilase family that performs thiol-assisted catalysis. Inhibition by SH reagents and the conserved cysteine residue in AguA and its homologues suggested that this enzyme is also involved in thiol-mediated catalysis.

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2003-03-01
2019-10-23
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References

  1. Baur, H., Luethi, E., Stalon, V., Mercenier, A. & Haas, D. ( 1989; ). Sequence analysis and expression of the arginine-deiminase and carbamate-kinase genes of Pseudomonas aeruginosa. Eur J Biochem 179, 53–60.[CrossRef]
    [Google Scholar]
  2. Bewley, M. C., Jeffrey, P. D., Patchett, M. L., Kanyo, Z. F. &, Baker. E. N. ( 1999; ). Crystal structure of Bacillus caldovelox arginase in complex with substrate and inhibitor reveals new insights into activation, inhibition and catalysis in the arginase family. Structure 7, 435–448.[CrossRef]
    [Google Scholar]
  3. Bork, F. &, Koonin. E. V. ( 1994; ). A new family of carbon-nitrogen hydrolases. Protein Sci 3, 1344–1346.[CrossRef]
    [Google Scholar]
  4. Chaudhuri, M. M. &, Ghosh. B. ( 1985; ). Agmatine deiminase in rice seedlings. Phytochemistry 24, 2433–2435.[CrossRef]
    [Google Scholar]
  5. Comai, L., Schilling-Cardoro, C., Mergia, A. & Houck, C. M. ( 1983; ). A new technique for genetic engineering of Agrobacterium Ti plasmid. Plasmid 10, 21–30.[CrossRef]
    [Google Scholar]
  6. Cunin, R., Glansdorff, N., Piérard, A. & Stalon, V. ( 1986; ). Biosynthesis and metabolism of arginine in bacteria. Microbiol Rev 50, 314–352.
    [Google Scholar]
  7. Dodson, G. &, Wlodawer. A. ( 1998; ). Catalytic triads and their relatives. Trends Biochem Sci 23, 347–352.[CrossRef]
    [Google Scholar]
  8. Fonzi, W. A. & Sypherd, P. S. ( 1987; ). The gene and the primary structure of ornithine decarboxylase from Saccharomyces cerevisiae. J Biol Chem 262, 10127–10133.
    [Google Scholar]
  9. Glansdorff, N. ( 1996; ). Biosynthesis of arginine and polyamines. In Escherichia coli and Salmonella: Cellular and Molecular Biology, 2nd edn, pp. 408–433. Edited by F. C. Neidhardt and others. Washington, DC: American Society for Microbiology.
  10. Haas, D. & Leisinger, T. ( 1975; ). N-acetylglutamate 5-phosphotransferase of Pseudomonas aeruginosa. Catalytic and regulatory properties. Eur J Biochem 52, 377–383.[CrossRef]
    [Google Scholar]
  11. Haas, D., Kurer, V. & Leisinger, T. ( 1972; ). N-acetylglutamate synthetase of Pseudomonas aeruginosa: an assay in vitro and feedback inhibition by arginine. Eur J Biochem 31, 290–295.[CrossRef]
    [Google Scholar]
  12. Haas, D., Matsumoto, H., Moretti, P., Stalon, V. & Mercenier, A. ( 1984; ). Arginine degradation in Pseudomonas aeruginosa mutants blocked in two-arginine catabolic pathways. Mol Gen Genet 193, 437–444.[CrossRef]
    [Google Scholar]
  13. Hoang, T. T., Karkhoff-Schweizer, R. R., Kutchma, A. J. & Schweizer, H. P. ( 1998; ). A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequence: application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene 212, 77–86.[CrossRef]
    [Google Scholar]
  14. Jann, A., Stalon, V., Vander Wauven, C., Leisinger, T. & Haas, D. ( 1986; ). N 2-Succinylated intermediates in arginine catabolic pathway of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 83, 4937–4941.[CrossRef]
    [Google Scholar]
  15. Kostyukova, A., Maeda, K., Krieger, I. & Maeda, Y. ( 2000; ). Domain structure of tropomodulin. Eur J Biochem 267, 6470–6475.[CrossRef]
    [Google Scholar]
  16. Löster, K., Baum, O., Hofmann, W. & Reutter, W. ( 1995; ). Chemical cross-linking leads to two high molecular mass aggregates of rat α1β1 integrin differing in their conformation but not in their composition. FEBS Lett 373, 234–238.[CrossRef]
    [Google Scholar]
  17. Lu, C.-D., Itoh, Y., Nakada, Y. & Jiang, Y. ( 2002; ). Functional analysis and regulation of the divergent spuABCDEFGH-spuI operons for polyamine uptake and utilization in Pseudomonas aeruginosa PAO1. J Bacteriol 184, 3765–3773.[CrossRef]
    [Google Scholar]
  18. McGraw, W. T., Potempa, J., Farley, D. & Travis, J. ( 1999; ). Purification, characterization, and sequence analysis of a potential virulence factor from Porphyromonas gingivalis. Peptidylarginine deiminase. Infect Immun 67, 3248–3256.
    [Google Scholar]
  19. McVey, C. E., Walsh, M. A., Dodson, G. G., Wilson, K. S. & Brannigan, J. A. ( 2001; ). Crystal structure of penicillin acylase enzyme–substrate complex: structural insights into the catalytic mechanism. J Mol Biol 313, 139–150.[CrossRef]
    [Google Scholar]
  20. Mercenier, A., Simon, J.-P., Haas, D. & Stalon, V. ( 1980; ). Catabolism of l-arginine by Pseudomonas aeruginosa. J Gen Appl Microbiol 116, 381–389.
    [Google Scholar]
  21. Morris, D. R. & Boeker, E. A. ( 1983; ). Biosynthetic and biodegradative ornithine and arginine decarboxylase from Escherichia coli. Methods Enzymol 94, 125–134.
    [Google Scholar]
  22. Nakada, Y. & Itoh, Y. ( 2002; ). Characterization and regulation of the gbuA gene, encoding guanidinobutyrase in the arginine dehydrogenase pathway of Pseudomonas aeruginosa PAO1. J Bacteriol 184, 3377–3384.[CrossRef]
    [Google Scholar]
  23. Nakada, Y., Jiang, Y., Nishijyo, T., Itoh, Y. & Lu, C.-D. ( 2001; ). Molecular characterization and regulation of the aguBA operon, responsible for agmatine utilization in Pseudomonas aeruginosa PAO1. J Bacteriol 183, 6517–6524.[CrossRef]
    [Google Scholar]
  24. Nishijyo, T., Haas, D. & Itoh, Y. ( 2001; ). The CbrA-CbrB two-component regulatory system controls the utilization of multiple carbon and nitrogen sources in Pseudomonas aeruginosa. Mol Microbiol 40, 917–931.[CrossRef]
    [Google Scholar]
  25. Pace, H. C. & Brenner, C. ( 2001; ). The nitrilase superfamily: classification, structure and function. Genome Biol 2, 1–8.
    [Google Scholar]
  26. Park, K. H. & Cho, Y. D. ( 1991; ). Purification of monomeric agmatine iminohydrolase from soybean. Biochem Biophys Res Commun 174, 32–36.[CrossRef]
    [Google Scholar]
  27. Perozich, J., Hempel, J. & Morris, S. M., Jr ( 1998; ). Roles of conserved residues in the arginase family. Biochim Biophys Acta 1382, 23–37.[CrossRef]
    [Google Scholar]
  28. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual. 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  29. Shin, S. S., Lee, T.-H., Ha, N.-C., Koo, H. M., Kim, S., Lee, H.-S., Kim, Y. S. & Oh, B.-H. ( 2002; ). Structure of malonamidase E2 reveals a novel Ser-cisSer-Lys catalytic triad in a new serine hydrolase fold that is prevalent in nature. EMBO J 21, 2509–2515.[CrossRef]
    [Google Scholar]
  30. Stover, C. K., Pham, X. Q., Erwin, A. L. & 28 other authors ( 2000; ). Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 406, 959–964.[CrossRef]
    [Google Scholar]
  31. Tabor, H. & Tabor, C. W. ( 1972; ). Biosynthesis and metabolism of 1,4-diaminobutane, spermidine, spermine, and related amines. Adv Enzymol Relat Areas Mol Biol 36, 203–268.
    [Google Scholar]
  32. Vieira, J. & Messing, J. ( 1987; ). Production of single-stranded plasmid DNA. Methods Enzymol 153, 3–11.
    [Google Scholar]
  33. Yanagisawa, H. & Suzuki, Y. ( 1981; ). Corn agmatine iminohydrolase: purification and properties. Plant Physiol 67, 697–700.[CrossRef]
    [Google Scholar]
  34. Yanagisawa, H. & Suzuki, Y. ( 1982; ). Purification and properties of N-carbamoylputrescine amidohydrolase from maize shoots. Phytochemistry 21, 2201–2203.[CrossRef]
    [Google Scholar]
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