Abstract
A two-dimensional electrophoretic method was used to show that protein degradation occurs immediately after the end of exponential growth but that its occurrence is masked in the usual assay methods for a 2-h period and that degradation is apparently nonselective with respect to protein molar mass or charge. The results suggest that considerable reutilization of internal amino acids may occur during sporulation regardless of the size of the external chase. Finally, the levels of intracellular proteinase activities present even at the end of exponential phase growth, as measuredin vitro, are sufficient to account for the maximum rates of protein degradation observedin vivo.
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References
Bernlohr R.W.: Role of amino acids in sporulation, pp. 75–87 inSpores III. Am.Soc.Microbiol., Ann Arbor 1965.
Bernlohr R.W.: Changes in amino acid permeation during sporulation.J. Bacteriol. 93, 1031–1044 (1967).
Bernlohr R.W.: Oxygen probes of protein turnover, amino acid transport, and protein synthesis inBacillus licheniformis.J. Biol.Chem. 247, 4893–4899 (1972).
Bernlohr R.W., Clark V.: Characterization and regulation of protease synthesis and activity inBacillus licheniformis.J. Bacteriol. 105, 276–283 (1971).
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. 72, 248–254 (1976).
Galandruccio J.H., Larrabee A.R.: Protein turnover in chemostat cultures ofEscherichia coli.Arch.Biochem. Biophys. 210, 270–274 (1981).
Chaloupka J., Philippová L., Čechová J., Strnadová M.: Relationship between intracellular proteolytic activity and protein turnover inBacillus megaterium.Acta Biol.Med.Germ. 36, 1515–1522 (1977).
Chaloupka J., Pillai K.R., Strnadová M.: The complex effect of temperature on protein degradation in sporulatingBacillus megaterium.FEMS Microbiol.Lett. 9, 107–110 (1980).
Chaloupka J., Strnadová M.: Degradation and replenishment of the labile protein fraction in non-growing cells of the asporogenic mutant ofBacillus megaterium.Folia Microbiol. 25, 191–200 (1980).
Davis-Mancini K., Lopez I.P., Hageman J.H.: Benzeneboronic acid selectively in hibits sporulation ofBacillus subtolis.J. Bacteriol. 136, 625–630 (1978).
Dougenik J.A., Sheehan D.E.: inSYMAP Reference Manual, Graduate School of Design, Harvard Boston 1977.
Eaton M.W., Ellar D.J.: Protein synthesis and breakdown in the mother-cell and forespore compartments during spore morphogenesis inBacillus megaterium.Biochem. J. 144, 327–337 (1974).
Goldberg A.L., St. John A.C.: Intracellular protein degradation in mammalian and bacterial cells.Ann.Rev.Biochem. 45, 747–803 (1976).
Goldman R.C.: Membrane protein alterations during the early stages of sporulation ofBacillus subtilis.J.Supramolec.Struct. 5, 457–473 (1976).
Hanson R.S., Blicharska J., Szulmajster J.: Relationship between the tricarboxylic acid cycle enzymes and sporulation inBacillus subtilis.Biochem.Biophys.Res.Comm. 17, 1–7 (1964).
Kerjan P., Keryer E., Szulmajster J.: Characterization of a thermosensitive sporulation mutant ofBacillus subtolis affected in the structural gene of an intracellular protease.Eur. J. Biochem. 98, 363–372 (1979).
Khan S.M., Darnall D.W.: The hydrolysis of 3-(2-furylacryoyl)-glycyl-l-leucine amide by thermolysin.Anal.Biochem. 86, 332–336 (1978).
Larrabee K.L., Phillips J.O., Williams G.J., Larrabee A.R.: The relative rates of protein synthesis and degradation in a growing culture ofEscherichia coli.J.Biol.Chem. 255, 4125–4130 (1980).
Linn T., Losick R.: The program of protein synthesis during sporulation inBacillus subtilis.Cell 8, 103–114 (1976).
Mandelstam J., Waites W.M.: Sporulation inBacillus subtilis. The role of exoprotease.Biochem. J. 109, 793–801 (1968).
Maurizi M.R., Brabson J.S., Switzer R.L.: Immunochemical studies of the inactivation of aspartate transcarbamylase by stationary phaseBacillus subtilis cells. Evidence for selective, energy-dependent degradation.J.Biol.Chem. 253, 5585–5593 (1978).
Maubizi M.R., Switzer R.L.: Proteolysis in bacterial sporulation.Curr.Topics Cell.Regul. 16, 164–224 (1980).
Mitani T., Kadota H.: Selectivity in protein degradation during sporulation ofBacillus subtilis.Agr.Biol.Chem. 40, 2319–2323 (1976).
Mosteller R.D., Goldstein R.V., Nishimoto K.R.: Metabolism of individual proteins in exponentially growingEscherichia coli.J.Biol.Chem. 255, 2524–2532 (1980).
Nakamura M., Okamura S., Izaki K., Takahashi H.: Further studies on an asporogenous mutant ofBacillus subtolis lackingin vivo proteolytic activity.J.Gen.Appl.Microbiol. 36, 119–132 (1980).
Nath K., Koch A.L.: Protein degradation inEscherichia coli. I. Measurement of rapidly and slowly decaying components.J.Biol.Chem. 245, 2889–2900 (1970).
Neame K.D., Homewood C.A.: pp. 115–123 inLiquid Scintillation Counting. John Wiley, New York 1974.
O’Farrell P.H.: High resolution two-dimensional electrophoresis of protein.J.Biol.Chem. 250, 4007–4021 (1975).
Ramaley R.F., Bueden L.: Replacement sporulation ofBacillus subtolis 168 in a chemically defined medium.J.Bacteriol. 101, 1–8 (1970).
Ruppen M.E., Switzeb R.L.: Degradation ofBacillus subtilis glutamine phosphoribosylpyrophosphate amidotransferasein vivo.J.Biol.Chem. 258, 2843–2851 (1983).
Sastry K.J., Srivistava O.P., Millet J., Fitzjames P.C., Aronson A.I.: Characterization ofBacillus subtilis mutants with a temperature-sensitive intracellular protease.J.Bacteriol. 153, 511–519 (1983).
Schroer D.W., St. John A.C.: Relative stability of membrane proteins inEscherichia coli. J.Bacteriol.146, 476–483 (1981).
Sekar V.:PhD Thesis. New Mexico State University, Las Cruces (USA) 1979.
Setlow P.: Protease and peptidase activities in growing and sporulating cells and dormant spores ofBacillus megaterium.J.Bacteriol. 122, 642–649 (1975).
Spizizen J.: Transformation of biochemically definedBacillus subtilis by deoxyribonucleate.Proc.Nat.Acad.Sci.USA 44, 1072–1078 (1958).
Spudich J.A., Kornberg A.: Biochemical studies of bacterial sporulation and germination. VII. Protein turnover during sporulation ofBacillus subtilis.J.Biol.Chem. 243, 4600–4605 (1968).
Vasantha N., Freese E.: Enzyme changes duringBacillus subtolis sporulation caused by deprivation of guanine nucleotides.J.Bacteriol. 144, 1119–1125 (1980).
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Sekar, V., Hageman, J.H. Protein turnover and proteolysis during sporulation ofBacillus subtilis . Folia Microbiol 32, 465–480 (1987). https://doi.org/10.1007/BF02877199
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DOI: https://doi.org/10.1007/BF02877199