The major native proteins of the leprosy bacillus.
This study addresses a major obstacle to vaccine development for leprosy, the isolation and characterization of the native protein antigens of the leprosy bacillus. Mycobacterium leprae harvested from armadillos was subjected to a simple fractionation protocol to arrive at the three major subcellular fractions, cell walls, cytoplasmic membrane, and soluble cytoplasm. The application of extensive detergent phase separations to membrane fractions allowed removal of lipoarabinomannan and the mannosyl phosphatidylinositols, and the recognition and purification of two major membrane proteins (MMP) of molecular mass 35 kDa (MMP-I) and 22 kDa (MMP-II); recovery of these proteins was about 0.5 mg each per g of M. leprae. MMP-I is N-blocked and is perhaps a lipoprotein. End group analysis on MMP-II indicates a new protein. Three major cytoplasmic proteins (MCP) of molecular mass 14 kDa (MCP-I), 17 kDa (MCP-II), and 28 kDa (MCP-III) were also recognized. MCP-I, the most abundant protein in M. leprae, represents 1% of the bacterial mass. End group analysis of the first 30 residues and immunoblotting studies demonstrate sizeable structural homology to a protein from Mycobacterium tuberculosis but immunological distinctiveness. MCP-I, which also occurs in highly immunogenic peptidoglycan-bound form, is a primary candidate for future vaccine development. The cell walls of M. leprae are also characterized by one major extractable protein, also of molecular mass 17 kDa. Thus the major antigens of the leprosy bacillus, protein and carbohydrate alike, are now nearer to complete definition.
Department of Microbiology, Colorado State University, Fort Collins 80523.
S W
SW
Hunter
B
B
Rivoire
V
V
Mehra
B R
BR
Bloom
P J
PJ
Brennan
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002458:Cell Fractionation; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009166:Mycobacterium leprae / Q000032:analysis*; D017830:Octoxynol; D011092:Polyethylene Glycols
D001426:Bacterial Proteins; D003902:Detergents; D008565:Membrane Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40