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. 1979 Mar;137(3):1324–1332. doi: 10.1128/jb.137.3.1324-1332.1979

Multiple Intracellular Peptidases in Neurospora crassa

Sai-Tee Tan 1,†, George A Marzluf 1
PMCID: PMC218316  PMID: 86534

Abstract

Neurospora crassa possesses multiple intracellular peptidases which display overlapping substrate specificities. They were readily detected by an in situ staining procedure for peptidases separated in polyacrylamide gels, within which the auxilliary enzyme, l-amino acid oxidase, was immobilized. Eleven different intracellular peptidases were identified by electrophoretic separation and verified by their individual patterns of substrate specificities. Most peptide substrates tested were hydrolyzed by several different peptidases. The multiple intracellular peptidases may play overlapping roles in several basic cell processes which involve peptidase activity. The amount of peptidase activity for leucylglycine present in crude extracts of cells grown under widely different conditions was relatively constant, suggesting that this enzyme may be constitutive, although alterations in the amounts of individual peptidase isozymes may occur. A single enzyme, designated peptidase II, was partially purified and obtained free from the other peptidase species. Peptidase II was found to be an aminopeptidase with activity toward many peptides of varied composition and size. It was more active with tripeptides than homologous dipeptides and showed strong activity toward methionine-containing peptides. This enzyme, with a molecular weight of about 37,000, was thermolabile at 65°C and was strongly inhibited by p-hydroxymercuribenzoate, Zn2+, Co2+, and Mn2+, but was insensitive to the serine protease inhibitor phenylmethylsulfonyl fluoride. Peptidase II apparently possesses an essential sulfhydryl group and may be a metalloenzyme.

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Selected References

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  1. Binkley F., Leibach F., King N. A new method of peptidase assay and the separation of three leucylglycinases of renal tissues. Arch Biochem Biophys. 1968 Nov;128(2):397–405. doi: 10.1016/0003-9861(68)90046-5. [DOI] [PubMed] [Google Scholar]
  2. Drucker H. Regulation of exocellular proteases in Neurospora crassa: induction and repression of enzyme synthesis. J Bacteriol. 1972 Jun;110(3):1041–1049. doi: 10.1128/jb.110.3.1041-1049.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hanson M. A., Marzluf G. A. Control of the synthesis of a single enzyme by multiple regulatory circuits in Neurospora crassa. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1240–1244. doi: 10.1073/pnas.72.4.1240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hanson M. A., Marzluf G. A. Regulation of a sulfur-controlled protease in Neurospora crassa. J Bacteriol. 1973 Nov;116(2):785–789. doi: 10.1128/jb.116.2.785-789.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Lewis W. H., Harris H. Human red cell peptidases. Nature. 1967 Jul 22;215(5099):351–355. doi: 10.1038/215351a0. [DOI] [PubMed] [Google Scholar]
  7. McHugh G. L., Miller C. G. Isolation and characterization of proline peptidase mutants of Salmonella typhimurium. J Bacteriol. 1974 Oct;120(1):364–371. doi: 10.1128/jb.120.1.364-371.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Miller C. G., Heiman C., Yen C. Mutants of Salmonella typhimurium deficient in an endoprotease. J Bacteriol. 1976 Jul;127(1):490–497. doi: 10.1128/jb.127.1.490-497.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Miller C. G., Mackinnon K. Peptidase mutants of Salmonella typhimurium. J Bacteriol. 1974 Oct;120(1):355–363. doi: 10.1128/jb.120.1.355-363.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Rice M. J. Supercontracting striated muscle in a vertebrate. Nature. 1973 May 25;243(5404):238–240. doi: 10.1038/243238a0. [DOI] [PubMed] [Google Scholar]
  11. Siepen D., Yu P. H., Kula M. R. Proteolytic enzymes of Neurospora crassa. Purification and some properties of five intracellular proteinases. Eur J Biochem. 1975 Aug 1;56(1):271–281. doi: 10.1111/j.1432-1033.1975.tb02230.x. [DOI] [PubMed] [Google Scholar]
  12. Smith A. E., Marcker K. A. Cytoplasmic methionine transfer RNAs from eukaryotes. Nature. 1970 May 16;226(5246):607–610. doi: 10.1038/226607a0. [DOI] [PubMed] [Google Scholar]
  13. THAYER P. S., HOROWITZ N. H. The L-amino acid oxidase of Neurospora. J Biol Chem. 1951 Oct;192(2):755–767. [PubMed] [Google Scholar]
  14. Vonder Haar R. A., Umbarger H. E. Isoleucine and valine metabolism in Escherichia coli. XIX. Inhibition of isoleucine biosynthesis by glycyl-leucine. J Bacteriol. 1972 Oct;112(1):142–147. doi: 10.1128/jb.112.1.142-147.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Wolfinbarger L., Jr, Marzluf G. A. Peptide utilization by amino acid auxotrophs of Neurospora crassa. J Bacteriol. 1974 Aug;119(2):371–378. doi: 10.1128/jb.119.2.371-378.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wolfinbarger L., Jr, Marzluf G. A. Size restriction on utilization of peptides by amino acid auxotrophs of Neurospora crassa. J Bacteriol. 1975 Jun;122(3):949–956. doi: 10.1128/jb.122.3.949-956.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wolfinbarger L., Jr, Marzluf G. A. Specificity and regulation of peptide transport on Neurospora crassa. Arch Biochem Biophys. 1975 Dec;171(2):637–644. doi: 10.1016/0003-9861(75)90074-0. [DOI] [PubMed] [Google Scholar]

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