Mapping and identification of the major cell wall-associated components of Mycobacterium leprae.
Mycobacterium leprae, an obligate intracellular pathogen, can be derived only from host tissue and thus affords the opportunity to study in vivo-expressed products responsible for the particular pathogenesis of leprosy. Despite considerable progress in the characterization of the proteins and secondary gene products of M. leprae, there is little information on the nature of the proteins associated with the cell envelope. M. leprae has been fractionated into its major subcellular components, cell wall, cytoplasmic membrane, and soluble cytosol. A number of biochemical markers, including diaminopimelic acid content, monosaccharide composition, mycolic acid, and glycolipid distribution, were applied to their characterization, and two-dimensional gel electrophoresis was used to map the component proteins. A total of 391 major proteins spots were resolved, and 8 proteins were identified based on their reactivity to a panel of monoclonal antibodies and/or relative pI size. Microsequencing of six protein spots present in the cell wall fraction allowed identification of new proteins, including the protein elongation factor EF-Tu and a homolog for the Mycobacterium tuberculosis MtrA response regulator. These results, together with previous studies, contribute to the progressive knowledge of the composition of the in vivo-expressed proteins of M. leprae.
Marques
M A
MA
Laboratório de Hanseníase, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, 21045-900 Rio de Janeiro, Brazil.
Chitale
S
S
Brennan
P J
PJ
Pessolani
M C
MC
D018528:ATP-Binding Cassette Transporters / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000302:isolation & purification; D001135:Armadillos; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002473:Cell Wall / Q000737:chemistry*; D002499:Centrifugation, Density Gradient; D018834:Chaperonin 60; D018833:Chaperonins / Q000302:isolation & purification; D003960:Diaminopimelic Acid / Q000032:analysis; D008055:Lipids / Q000032:analysis; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis; D009166:Mycobacterium leprae / Q000737:chemistry*; D010444:Peptide Elongation Factor Tu / Q000032:analysis; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000737:chemistry
D018528:ATP-Binding Cassette Transporters; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D018834:Chaperonin 60; D008055:Lipids; D009005:Monosaccharides; C100938:MtrA protein, Bacteria; C074119:heat-shock protein 65, Mycobacterium; D003960:Diaminopimelic Acid; D018833:Chaperonins; D010444:Peptide Elongation Factor Tu