Home LiteratureArticle Details
PMID: 1563797 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Characterization of the major membrane protein of virulent Mycobacterium tuberculosis.

Infection and immunity ·Vol. 60 ·No. 5 ·1992-05-00 ·Pages 2066-74

Lee BY, Hefta SA, Brennan PJ

Abstract

A protein with a molecular mass of 19 kDa was isolated and purified from enriched membrane fractions of the virulent Erdman strain of Mycobacterium tuberculosis. The protein is different from another 19-kDa protein, a lipoprotein, that was recently described (D. B. Young and T. R. Garbe, Res. Microbiol. 142:55-65, 1991). The sequencing strategy applied to this major membrane protein employed four different endoproteinases and resulted in sufficient overlapping peptide sequences for assignment of the entire protein sequence. Electron spray ionization mass spectrometry demonstrated a measured mass of 16,100, deviating from the predicted mass by only 2.86 atomic mass units. The sequence of this protein is unique. However, some similarities with other low-molecular-weight heat shock proteins were observed. Immunoblotting indicated that this protein is highly expressed in the virulent strains of M. tuberculosis. Its application to sera from patients with pulmonary tuberculosis showed promise as a serodiagnostic tool.

MeSH Terms
Amino Acid Sequence Antibodies, Bacterial/analysis Bacterial Proteins/analysis,chemistry,isolation & purification Humans Mass Spectrometry Membrane Proteins/analysis,chemistry,isolation & purification Molecular Sequence Data Molecular Weight Mycobacterium tuberculosis/chemistry,pathogenicity Virulence
Chemicals
Antibodies, Bacterial Bacterial Proteins Membrane Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lee B Y
Department of Microbiology, Colorado State University, Fort Collins 80523.
Hefta S A
Brennan P J
References (40)
40 references, click to expand
  1. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity.
    Anal Biochem. 1981 Nov 1;117(2):307-10 PMID: 6172996
  2. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  3. An analysis of mRNAs for a group of heat shock proteins of soybean using cloned cDNAs.
    J Mol Appl Genet. 1982;1(4):301-14 PMID: 6896722
  4. Structural studies on the myo-inositol phospholipids of Mycobacterium tuberculosis (var. bovis, strain BCG).
    J Biol Chem. 1963 Jan;238:69-76 PMID: 13966180
  5. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  6. The 14,000-molecular-weight antigen of Mycobacterium tuberculosis is related to the alpha-crystallin family of low-molecular-weight heat shock proteins.
    J Bacteriol. 1992 Feb;174(4):1352-9 PMID: 1370952
  7. Mycobacterial protein antigens: a compilation.
    Mol Microbiol. 1992 Jan;6(2):133-45 PMID: 1545700
  8. The universally conserved GroE (Hsp60) chaperonins.
    Annu Rev Microbiol. 1991;45:301-25 PMID: 1683763
  9. Chemical synthesis and seroreactivity of a neoantigen containing the oligosaccharide hapten of the Mycobacterium tuberculosis-specific phenolic glycolipid.
    J Infect Dis. 1991 Jan;163(1):161-8 PMID: 1898609
  10. Enzymatic and chemical digestion of proteins for mass spectrometry.
    Methods Enzymol. 1990;193:361-74 PMID: 1963671
  11. Peptidoglycan-associated polypeptides of Mycobacterium tuberculosis.
    J Bacteriol. 1990 Feb;172(2):1005-13 PMID: 2105289
  12. Mycobacterial antigens: a review of their isolation, chemistry, and immunological properties.
    Microbiol Rev. 1978 Mar;42(1):84-113 PMID: 88663
  13. Site of inhibitory action of isoniazid in the synthesis of mycolic acids in Mycobacterium tuberculosis.
    J Lipid Res. 1975 Jul;16(4):308-17 PMID: 806645
  14. Heat shock proteins and antigens of Mycobacterium tuberculosis.
    Infect Immun. 1991 Sep;59(9):3086-93 PMID: 1679042
  15. Lipoarabinomannan, a possible virulence factor involved in persistence of Mycobacterium tuberculosis within macrophages.
    Infect Immun. 1991 May;59(5):1755-61 PMID: 1850379
  16. Lipoprotein antigens of Mycobacterium tuberculosis.
    Res Microbiol. 1991 Jan;142(1):55-65 PMID: 1906192
  17. Evidence for the presence of a phosphatidylinositol anchor on the lipoarabinomannan and lipomannan of Mycobacterium tuberculosis.
    J Biol Chem. 1990 Jun 5;265(16):9272-9 PMID: 2111816
  18. The major native proteins of the leprosy bacillus.
    J Biol Chem. 1990 Aug 25;265(24):14065-8 PMID: 2201679
  19. The heat-shock response.
    Annu Rev Biochem. 1986;55:1151-91 PMID: 2427013
  20. Tuberculosis and human immunodeficiency virus infection.
    J Infect Dis. 1989 Jan;159(1):96-100 PMID: 2642524
  21. Major integral membrane protein immunogens of Treponema pallidum are proteolipids.
    Infect Immun. 1989 Sep;57(9):2872-7 PMID: 2668191
  22. Microsequence analysis of peptides and proteins. VI. A continuous flow reactor for sample concentration and sequence analysis.
    Anal Biochem. 1987 Jun;163(2):517-29 PMID: 3662001
  23. Genes for low-molecular-weight heat shock proteins of soybeans: sequence analysis of a multigene family.
    Mol Cell Biol. 1985 Dec;5(12):3417-28 PMID: 3939317
  24. Separation of the inner (cytoplasmic) and outer membranes of Gram-negative bacteria.
    Methods Enzymol. 1974;31:642-53 PMID: 4608978
  25. Tuberculosis in developing countries: burden, intervention and cost.
    Bull Int Union Tuberc Lung Dis. 1990 Mar;65(1):6-24 PMID: 2190653
  26. Immunogenic integral membrane proteins of Borrelia burgdorferi are lipoproteins.
    Infect Immun. 1990 Apr;58(4):983-91 PMID: 2318538
  27. The specific 18-kilodalton antigen of Mycobacterium leprae is present in Mycobacterium habana and functions as a heat-shock protein.
    J Immunol. 1990 Mar 1;144(5):1922-5 PMID: 2407785
  28. Antigenic proteins of Mycobacterium leprae. Complete sequence of the gene for the 18-kDa protein.
    J Immunol. 1988 Jan 15;140(2):597-601 PMID: 2447183
  29. Tuberculosis in the United States.
    JAMA. 1989 Jul 21;262(3):385-9 PMID: 2661873
  30. A protein antigen of Mycobacterium leprae is related to a family of small heat shock proteins.
    J Bacteriol. 1988 Dec;170(12):5919-21 PMID: 3056923
  31. The antigens of Mycobacterium tuberculosis, H37Rv, studied by crossed immunoelectrophoresis. Comparison with a reference system for Mycobacterium bovis, BCG.
    Scand J Immunol. 1988 Feb;27(2):223-39 PMID: 3124264
  32. Major membrane surface proteins of Mycoplasma hyopneumoniae selectively modified by covalently bound lipid.
    J Bacteriol. 1987 Dec;169(12):5546-55 PMID: 3680170
  33. Results of a World Health Organization-sponsored workshop to characterize antigens recognized by mycobacterium-specific monoclonal antibodies.
    Infect Immun. 1986 Feb;51(2):718-20 PMID: 3943911
  34. Microsequence analysis of peptides and proteins. V. Design and performance of a novel gas-liquid-solid phase instrument.
    Anal Biochem. 1985 Jun;147(2):315-30 PMID: 4014677
  35. Antigenic diversity of Mycobacterium tuberculosis and Mycobacterium bovis detected by means of monoclonal antibodies.
    Lancet. 1981 Jul 25;2(8239):167-9 PMID: 6166820
  36. Characteristics of n-octyl beta-D-thioglucopyranoside, a new non-ionic detergent useful for membrane biochemistry.
    Biochem J. 1984 Sep 15;222(3):829-32 PMID: 6237644
  37. Phase separation of integral membrane proteins in Triton X-114 solution.
    J Biol Chem. 1981 Feb 25;256(4):1604-7 PMID: 6257680
  38. Production and characterization of monoclonal antibodies to Mycobacterium tuberculosis, M. bovis (BCG) and M. leprae.
    Clin Exp Immunol. 1984 Dec;58(3):511-21 PMID: 6439449
  39. A simple and rapid purification of commercial trypsin and chymotrypsin by reverse-phase high-performance liquid chromatography.
    Anal Biochem. 1982 Jul 1;123(2):408-12 PMID: 7125214
  40. Proteins of the outer membrane of gram-negative bacteria.
    Annu Rev Microbiol. 1980;34:369-422 PMID: 6254441
Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.60.5.2066-2074.1992
Published
1992-05-00
Pages
2066-74
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC257116
Subset
IM
Grants
NIAID NIH HHS · AI-27288 · United States
NCI NIH HHS · CA-33572 · United States
External Links
PubMed source DOI: 10.1128/iai.60.5.2066-2074.1992 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2