Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Apr;62(4):1150–1155. doi: 10.1128/iai.62.4.1150-1155.1994

Purification and characterization of a high-molecular-weight outer membrane protein of Moraxella (Branhamella) catarrhalis.

K L Klingman 1, T F Murphy 1
PMCID: PMC186244  PMID: 8132320

Abstract

Moraxella (Branhamella) catarrhalis is an important bacterial cause of otitis media in children and lower respiratory tract infections in adults. In this study, we describe the presence of a novel high-molecular-weight outer membrane protein (HMW-OMP). This protein varies from 350 to 720 kDa in apparent molecular mass among strains by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The protein was detected on SDS-PAGE in 13 of 14 strains tested. We developed a monoclonal antibody and polyclonal antisera to this protein. In immunoblot assays, the protein was present in all 14 strains tested. The immunoblot assays suggest that the protein has at least one epitope that is conserved among strains. A purification method using anion-exchange chromatography is described. Treatment of outer membrane preparations and purified protein by heat and reducing agents did not change the apparent molecular mass of the HMW-OMP. Formic acid treatment of outer membrane preparations and purified HMW-OMP produced a single band with an apparent molecular mass of 120 to 140 kDa. We postulate that this may be the monomer of an oligomeric protein. The HMW-OMP, which varies in molecular mass among strains and is antigenically conserved, will be studied further to determine its role in the human immune response and may be useful as a marker in studying strain acquisition in patients.

Full text

PDF
1150

Images in this article

1152Image
on p.1152
1152Image
on p.1152
1153Image
on p.1153
1153Image
on p.1153
1153Image
on p.1153
1154Image
on p.1154

Selected References

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

  1. Catlin B. W. Branhamella catarrhalis: an organism gaining respect as a pathogen. Clin Microbiol Rev. 1990 Oct;3(4):293–320. doi: 10.1128/cmr.3.4.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Collinson S. K., Emödy L., Müller K. H., Trust T. J., Kay W. W. Purification and characterization of thin, aggregative fimbriae from Salmonella enteritidis. J Bacteriol. 1991 Aug;173(15):4773–4781. doi: 10.1128/jb.173.15.4773-4781.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Doern G. V. Branhamella catarrhalis--an emerging human pathogen. Diagn Microbiol Infect Dis. 1986 Mar;4(3):191–201. doi: 10.1016/0732-8893(86)90098-2. [DOI] [PubMed] [Google Scholar]
  4. Hager H., Verghese A., Alvarez S., Berk S. L. Branhamella catarrhalis respiratory infections. Rev Infect Dis. 1987 Nov-Dec;9(6):1140–1149. doi: 10.1093/clinids/9.6.1140. [DOI] [PubMed] [Google Scholar]
  5. Hansen M. V., Wilde C. E., 3rd Conservation of peptide structure of outer membrane protein-macromolecular complex from Neisseria gonorrhoeae. Infect Immun. 1984 Mar;43(3):839–845. doi: 10.1128/iai.43.3.839-845.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hockfield S. A Mab to a unique cerebellar neuron generated by immunosuppression and rapid immunization. Science. 1987 Jul 3;237(4810):67–70. doi: 10.1126/science.3603010. [DOI] [PubMed] [Google Scholar]
  7. Judd R. C. Peptide mapping. Methods Enzymol. 1990;182:613–626. doi: 10.1016/0076-6879(90)82048-7. [DOI] [PubMed] [Google Scholar]
  8. Kennett R. H. Cell fusion. Methods Enzymol. 1979;58:345–359. doi: 10.1016/s0076-6879(79)58149-x. [DOI] [PubMed] [Google Scholar]
  9. Klingman K. L., Jansen E. M., Murphy T. F. Nearest neighbor analysis of outer membrane proteins of nontypeable Haemophilus influenzae. Infect Immun. 1988 Dec;56(12):3058–3063. doi: 10.1128/iai.56.12.3058-3063.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Murphy T. F., Bartos L. C. Surface-exposed and antigenically conserved determinants of outer membrane proteins of Branhamella catarrhalis. Infect Immun. 1989 Oct;57(10):2938–2941. doi: 10.1128/iai.57.10.2938-2941.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Murphy T. F., Loeb M. R. Isolation of the outer membrane of Branhamella catarrhalis. Microb Pathog. 1989 Mar;6(3):159–174. doi: 10.1016/0882-4010(89)90066-1. [DOI] [PubMed] [Google Scholar]
  12. Murphy T. F. Studies of the outer membrane proteins of Branhamella catarrhalis. Am J Med. 1990 May 14;88(5A):41S–45S. doi: 10.1016/0002-9343(90)90261-b. [DOI] [PubMed] [Google Scholar]
  13. Newhall W. J., Wilde C. E., 3rd, Sawyer W. D., Haak R. A. High-molecular-weight antigenic protein complex in the outer membrane of Neisseria gonorrhoeae. Infect Immun. 1980 Feb;27(2):475–482. doi: 10.1128/iai.27.2.475-482.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nicotra B., Rivera M., Luman J. I., Wallace R. J., Jr Branhamella catarrhalis as a lower respiratory tract pathogen in patients with chronic lung disease. Arch Intern Med. 1986 May;146(5):890–893. [PubMed] [Google Scholar]
  15. Olmsted J. B. Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem. 1981 Dec 10;256(23):11955–11957. [PubMed] [Google Scholar]
  16. Sarwar J., Campagnari A. A., Kirkham C., Murphy T. F. Characterization of an antigenically conserved heat-modifiable major outer membrane protein of Branhamella catarrhalis. Infect Immun. 1992 Mar;60(3):804–809. doi: 10.1128/iai.60.3.804-809.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  18. Tsai W. M., Larsen S. H., Wilde C. E., 3rd Cloning and DNA sequence of the omc gene encoding the outer membrane protein-macromolecular complex from Neisseria gonorrhoeae. Infect Immun. 1989 Sep;57(9):2653–2659. doi: 10.1128/iai.57.9.2653-2659.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Van Hare G. F., Shurin P. A., Marchant C. D., Cartelli N. A., Johnson C. E., Fulton D., Carlin S., Kim C. H. Acute otitis media caused by Branhamella catarrhalis: biology and therapy. Rev Infect Dis. 1987 Jan-Feb;9(1):16–27. doi: 10.1093/clinids/9.1.16. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES