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. 1978 Dec;75(12):5998–6001. doi: 10.1073/pnas.75.12.5998

Escherichia coli mutants defective in dipeptidyl carboxypeptidase.

C E Deutch, R L Soffer
PMCID: PMC393104  PMID: 216006

Abstract

Two independent mutants of Escherichia coli deficient in dipeptidyl carboxypeptidase activity (Dep-) were isolated after mutagenesis with ethyl methanesulfonate. Mating experiments and introduction of specific episomes indicated that the responsible gene was located at approximately 27--31 min on the E. coli chromosome. The Dep- mutants differed from the parental strain in their inability to grow with N-acetylalanylalanylalanine as the sole nitrogen source. Revertants selected for growth on this substrate of the enzyme were found to have reacquired the activity. Enzyme activity was highly sensitive to inhibition by 1-(D-3-mercapto-2-methylpropanoyl)-L-proline (SQ 14225), a potent inhibitor of mammalian dipeptidyl carboxypeptidase (angiotensin-converting enzyme, peptidyl dipeptidase, EC 3.4.15.1). This compound also reduced the rate of growth of the wild type with N-acetylalanylalanylalanine but not with ammonium sulfate. A fraction of the enzyme was released into the medium by osmotic shock, indicating that its presence in the periplasmic space may account for growth with N-acetylated peptides that cannot be taken up by E. coli. In addition to providing information about the specific role of this exopeptidase in E. coli, the Dep- mutants may prove useful for delineating the regulation and cellular function of dipeptidyl carboxypeptidases in higher organisms.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Case D. B., Wallace J. M., Keim H. J., Weber M. A., Sealey J. E., Laragh J. H. Possible role of renin in hypertension as suggested by renin-sodium profiling and inhibition of converting enzyme. N Engl J Med. 1977 Mar 24;296(12):641–646. doi: 10.1056/NEJM197703242961201. [DOI] [PubMed] [Google Scholar]
  2. Cheung H. S., Cushman D. W. Inhibition of homogeneous angiotensin-converting enzyme of rabbit lung by synthetic venom peptides of Bothrops jararaca. Biochim Biophys Acta. 1973 Feb 15;293(2):451–463. doi: 10.1016/0005-2744(73)90352-5. [DOI] [PubMed] [Google Scholar]
  3. Cushman D. W., Cheung H. S., Sabo E. F., Ondetti M. A. Design of potent competitive inhibitors of angiotensin-converting enzyme. Carboxyalkanoyl and mercaptoalkanoyl amino acids. Biochemistry. 1977 Dec 13;16(25):5484–5491. doi: 10.1021/bi00644a014. [DOI] [PubMed] [Google Scholar]
  4. Das M., Soffer R. L. Pulmonary angiotensin-converting enzyme antienzyme antibody. Biochemistry. 1976 Nov 16;15(23):5088–5094. doi: 10.1021/bi00668a022. [DOI] [PubMed] [Google Scholar]
  5. Das M., Soffer R. L. Pulmonary angiotensin-converting enzyme. Structural and catalytic properties. J Biol Chem. 1975 Sep 10;250(17):6762–6768. [PubMed] [Google Scholar]
  6. GILVARG C., KATCHALSKI E. PEPTIDE UTILIZATION IN ESCHERICHIA COLI. J Biol Chem. 1965 Jul;240:3093–3098. [PubMed] [Google Scholar]
  7. Gavras H., Brunner H. R., Turini G. A., Kershaw G. R., Tifft C. P., Cuttelod S., Gavras I., Vukovich R. A., McKinstry D. N. Antihypertensive effect of the oral angiotensin converting-enzyme inhibitor SQ 14225 in man. N Engl J Med. 1978 May 4;298(18):991–995. doi: 10.1056/NEJM197805042981803. [DOI] [PubMed] [Google Scholar]
  8. HERZENBERG L. A. Studies on the induction of beta-galactosidase in a cryptic strain of Escherichia coli. Biochim Biophys Acta. 1959 Feb;31(2):525–538. doi: 10.1016/0006-3002(59)90029-0. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Low K. B. Escherichia coli K-12 F-prime factors, old and new. Bacteriol Rev. 1972 Dec;36(4):587–607. doi: 10.1128/br.36.4.587-607.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Miller R. E., Stadtman E. R. Glutamate synthase from Escherichia coli. An iron-sulfide flavoprotein. J Biol Chem. 1972 Nov 25;247(22):7407–7419. [PubMed] [Google Scholar]
  12. Neu H. C., Heppel L. A. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem. 1965 Sep;240(9):3685–3692. [PubMed] [Google Scholar]
  13. Nossal N. G., Heppel L. A. The release of enzymes by osmotic shock from Escherichia coli in exponential phase. J Biol Chem. 1966 Jul 10;241(13):3055–3062. [PubMed] [Google Scholar]
  14. Ondetti M. A., Rubin B., Cushman D. W. Design of specific inhibitors of angiotensin-converting enzyme: new class of orally active antihypertensive agents. Science. 1977 Apr 22;196(4288):441–444. doi: 10.1126/science.191908. [DOI] [PubMed] [Google Scholar]
  15. Ondetti M. A., Williams N. J., Sabo E. F., Pluscec J., Weaver E. R., Kocy O. Angiotensin-converting enzyme inhibitors from the venom of Bothrops jararaca. Isolation, elucidation of structure, and synthesis. Biochemistry. 1971 Oct 26;10(22):4033–4039. doi: 10.1021/bi00798a004. [DOI] [PubMed] [Google Scholar]
  16. Payne J. W. The requirement for the protonated -amino group for the transport of peptides in Escherichia coli. Biochem J. 1971 Jun;123(2):245–253. doi: 10.1042/bj1230245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Soffer R. L. Angiotensin-converting enzyme and the regulation of vasoactive peptides. Annu Rev Biochem. 1976;45:73–94. doi: 10.1146/annurev.bi.45.070176.000445. [DOI] [PubMed] [Google Scholar]
  18. Yaron A. Dipeptidyl carboxypeptidase from Escherichia coli. Methods Enzymol. 1976;45:599–610. doi: 10.1016/s0076-6879(76)45053-x. [DOI] [PubMed] [Google Scholar]
  19. Yaron A., Mlynar D., Berger A. A dipeptidocarboxypeptidase from E. coli. Biochem Biophys Res Commun. 1972 May 26;47(4):897–902. doi: 10.1016/0006-291x(72)90577-3. [DOI] [PubMed] [Google Scholar]

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