Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Apr;150(1):303–311. doi: 10.1128/jb.150.1.303-311.1982

Bacillus megaterium spore protease: purification, radioimmunoassay, and analysis of antigen level and localization during growth, sporulation, and spore germination.

C A Loshon, P Setlow
PMCID: PMC220114  PMID: 6801023

Abstract

The protease which initiates the massive protein degradation early in bacterial spore germination has been purified from Bacillus megaterium spores. The enzyme has a molecular weight of 160,000 and contains four apparently identical subunits, but only the tetramer is enzymatically active. A radioimmunoassay has been developed for this enzyme and has been used to show that the protease is absent from growing cells, but appears early in sporulation within the developing forespore. In contrast, the protease antigen disappears rapidly during spore germination, in parallel with the loss in enzyme activity.

Full text

PDF
306

Images in this article

306Image
on p.306

Selected References

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

  1. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. CHURCH B. D., HALVORSON H. Intermediate metabolism of aerobic spores. I. Activation of glucose oxidation in spores of Bacillus cereus var terminalis. J Bacteriol. 1957 Apr;73(4):470–476. doi: 10.1128/jb.73.4.470-476.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Davies G. E., Stark G. R. Use of dimethyl suberimidate, a cross-linking reagent, in studying the subunit structure of oligomeric proteins. Proc Natl Acad Sci U S A. 1970 Jul;66(3):651–656. doi: 10.1073/pnas.66.3.651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dignam S. S., Setlow P. Bacillus megaterium spore protease. Action of the enzyme on peptides containing the amino acid sequence cleaved by the enzyme in vivo. J Biol Chem. 1980 Sep 25;255(18):8408–8412. [PubMed] [Google Scholar]
  6. 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]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  9. Postemsky C. J., Dignam S. S., Setlow P. Isolation and characterization of Bacillus megaterium mutants containing decreased levels of spore protease. J Bacteriol. 1978 Sep;135(3):841–850. doi: 10.1128/jb.135.3.841-850.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Setlow B., Setlow P. Localization of low-molecular-weight basic proteins in Bacillus megaterium spores by cross-linking with ultraviolet light. J Bacteriol. 1979 Aug;139(2):486–494. doi: 10.1128/jb.139.2.486-494.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Setlow P. Identification and localization of the major proteins degraded during germination of Bacillus megaterium spores. J Biol Chem. 1975 Oct 25;250(20):8159–8167. [PubMed] [Google Scholar]
  12. Setlow P., Kornberg A. Biochemical studies of bacterial sporulation and germination. XVII. Sulfhydryl and disulfide levels in dormancy and germination. J Bacteriol. 1969 Dec;100(3):1155–1160. doi: 10.1128/jb.100.3.1155-1160.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Setlow P., Primus G. Protein metabolism during germination of Bacillus megaterium spores. I. Protein synthesis and amino acid metabolism. J Biol Chem. 1975 Jan 25;250(2):623–630. [PubMed] [Google Scholar]
  14. Setlow P. Purification and properties of a specific proteolytic enzyme present in spores of Bacillus magaterium. J Biol Chem. 1976 Dec 25;251(24):7853–7862. [PubMed] [Google Scholar]
  15. Siegel L. M., Monty K. J. Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. Biochim Biophys Acta. 1966 Feb 7;112(2):346–362. doi: 10.1016/0926-6585(66)90333-5. [DOI] [PubMed] [Google Scholar]
  16. Singh R. P., Setlow B., Setlow P. Levels of small molecules and enzymes in the mother cell compartment and the forespore of sporulating Bacillus megaterium. J Bacteriol. 1977 Jun;130(3):1130–1138. doi: 10.1128/jb.130.3.1130-1138.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES