Search for the molecular basis of morphological variation in Mycobacterium avium.
Isolates of Mycobacterium avium exhibit three different colonial variations: smooth domed (SmD), smooth transparent (SmT), and rough (Rg). Because the discrimination between morphotypes is founded on morphological rather than molecular principles and because of the absence of consensus over the relevance of morphology to pathogenesis and drug sensitivity, a comparative study at the protein level was undertaken. By direct immunization of BALB/c mice with the soluble sonicate of one of the morphotypes of M. avium serovar 2, eight monoclonal antibodies (MAbs) were identified, of which one was M. avium specific. Cross immunization of syngeneic mice with serum-absorbed antigens allowed the generation of 15 further MAbs; 11 were M. avium or M. avium complex specific, but none of them was morphotype specific. Subcellular fractions analyzed by electrophoresis showed similar profiles, with the exception of a cytosolic protein with a relative molecular mass of ca. 66 kDa (protein SmT 66), which was most highly expressed in SmT variants of M. avium serotypes 2 and 4. Because a well-known, ubiquitous stress-heat shock protein (hsp65) has a similar molecular mass, protein SmT 66 was compared with hsp65. Western blot (immunoblot) analyses using several cross-reacting MAbs and N-terminal amino acid sequencing established that this protein was not the ubiquitous stress protein. Thus, SmT 66 is the first product to be described which might be associated with the SmT morphotype.
Prinzis
S
S
Department of Microbiology, Colorado State University, Fort Collins 80523.
Rivoire
B
B
Brennan
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000032:analysis*; D006360:Heat-Shock Proteins / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009162:Mycobacterium avium / Q000737:chemistry* / Q000166:cytology
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D006360:Heat-Shock Proteins