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. 1997 Oct;63(10):3770–3775. doi: 10.1128/aem.63.10.3770-3775.1997

Further characterization of Renibacterium salmoninarum extracellular products.

T A Barton 1, L A Bannister 1, S G Griffiths 1, W H Lynch 1
PMCID: PMC168686  PMID: 9480644

Abstract

Renibacterium salmoninarum, the agent of bacterial kidney disease in salmonids, releases high concentrations of extracellular protein in tissues of infected fish. The extracellular protein consists almost entirely of a 57-kDa protein and derivatives of degradation and aggregation of the same molecule. The 57-kDa protein and its derivatives were fractionated into defined ranges of molecular mass. Separated fractions continued to produce degradation and aggregation products. One-dimensional electrophoretic separation of extracellular protein revealed a number of proteolytically active bands from > 100 to approximately 18 kDa associated with various 57-kDa protein derivatives in the different molecular mass fractions. Two-dimensional separation of extracellular protein showed that continued degradation and aggregation, similar both in location and behavior to some of the 57-kDa protein derivatives, was also displayed by the proteolytically active bands after their separation. Effects of reducing agents and sulfhydryl group proteinase inhibitors indicated a common mechanism for the proteolytically active polypeptides characteristic of a thiol proteinase. The results suggested that the 57-kDa protein and some of its derivatives undergo autolytic cleavage, releasing a proteolytically active polypeptide(s) of at least 18 kDa. Soluble polysaccharide-like material also was detected in extracellular products and tissue from infected fish. Antiserum to the polysaccharide-like material cross-reacted with O-polysaccharide of the fish pathogen Aeromonas salmonicida, suggesting some structural similarity between these polysaccharides. The polysaccharide and the proteolytic activity associated with the 57-kDa protein derivatives should be investigated with respect to the pathogenesis of R. salmoninarum infections.

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Selected References

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  1. Daly J. G., Stevenson R. M. Characterization of the Renibacterium salmoninarum haemagglutinin. J Gen Microbiol. 1990 May;136(5):949–953. doi: 10.1099/00221287-136-5-949. [DOI] [PubMed] [Google Scholar]
  2. Dubreuil D., Lallier R., Jacques M. Immunoelectron microscopic demonstration that Renibacterium salmoninarum is encapsulated. FEMS Microbiol Lett. 1990 Jan 1;54(1-3):313–315. doi: 10.1016/0378-1097(90)90304-9. [DOI] [PubMed] [Google Scholar]
  3. Dubreuil J. D., Jacques M., Graham L., Lallier R. Purification, and biochemical and structural characterization of a fimbrial haemagglutinin of Renibacterium salmoninarum. J Gen Microbiol. 1990 Dec;136(12):2443–2448. doi: 10.1099/00221287-136-12-2443. [DOI] [PubMed] [Google Scholar]
  4. Griffiths S. G., Lynch W. H. Characterization of Aeromonas salmonicida mutants with low-level resistance to multiple antibiotics. Antimicrob Agents Chemother. 1989 Jan;33(1):19–26. doi: 10.1128/aac.33.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Griffiths S. G., Lynch W. H. Characterization of Aeromonas salmonicida variants with altered cell surfaces and their use in studying surface protein assembly. Arch Microbiol. 1990;154(3):308–312. doi: 10.1007/BF00248973. [DOI] [PubMed] [Google Scholar]
  6. Jacobs S. Effect of isoelectric focusing on the amino-acid composition of proteins. Analyst. 1973 Jan;98(162):25–33. doi: 10.1039/an9739800025. [DOI] [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. Pelkonen S., Finne J. Polyacrylamide gel electrophoresis of capsular polysaccharides of bacteria. Methods Enzymol. 1989;179:104–110. doi: 10.1016/0076-6879(89)79118-7. [DOI] [PubMed] [Google Scholar]
  9. Rawlings N. D., Barrett A. J. Families of cysteine peptidases. Methods Enzymol. 1994;244:461–486. doi: 10.1016/0076-6879(94)44034-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Rockey D. D., Turaga P. S., Wiens G. D., Cook B. A., Kaattari S. L. Serine proteinase of Renibacterium salmoninarum digests a major autologous extracellular and cell-surface protein. Can J Microbiol. 1991 Oct;37(10):758–763. doi: 10.1139/m91-130. [DOI] [PubMed] [Google Scholar]
  11. Szewczyk B., Summers D. F. Preparative elution of proteins blotted to Immobilon membranes. Anal Biochem. 1988 Jan;168(1):48–53. doi: 10.1016/0003-2697(88)90008-5. [DOI] [PubMed] [Google Scholar]
  12. Wiens G. D., Kaattari S. L. Monoclonal antibody characterization of a leukoagglutinin produced by Renibacterium salmoninarum. Infect Immun. 1991 Feb;59(2):631–637. doi: 10.1128/iai.59.2.631-637.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wood S. C., McCashion R. N., Lynch W. H. Multiple low-level antibiotic resistance in Aeromonas salmonicida. Antimicrob Agents Chemother. 1986 Jun;29(6):992–996. doi: 10.1128/aac.29.6.992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Woodward M. P., Young W. W., Jr, Bloodgood R. A. Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. J Immunol Methods. 1985 Apr 8;78(1):143–153. doi: 10.1016/0022-1759(85)90337-0. [DOI] [PubMed] [Google Scholar]

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