Amphiphilic properties of spiralin, the major surface antigen of spiroplasmas. A preliminary report.
Spiralin, a 28-kDa (kilodalton) polypeptide, is the major antigen of Spiroplasma citri and S. melliferum in which it usually represents more than 20% of total membrane protein. The amino acid compositions of the spiralins purified from both spiroplasma species unambiguously show that these proteins are homologous. In addition, several lines of evidence indicate that such a protein is present in the menbrane of S. apis. A 25-kDa polypeptide antigenically related to S. citri spiralin has also been purified from the membrane of the nonhelical variant ASP-1. The spiralin of S. melliferum B88 has been used as a model for extensive characterization. This antigen binds detergent under nondenaturing conditions, can be incorporated into liposomes, and forms protein micelles upon gentle removal of detergent. Digestion of the micelles with trypsin leads to the precipitation of an insoluble material containing a major polypeptide of 3.9 kDa. The amino acid composition of this fragment is different from that of intact spiralin. It is highly enriched in glycine and serine and, as an insoluble peptide, exhibits an unexpectedly high polarity index (PI = 51.4%). Screening for acyl proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunodetection in the membrane of S. melliferum indicates that spiralin is actually acylated. This set of properties is evidence that spiralin is an intrinsic membrane protein and strongly suggests that acylation triggers or facilitates integration of the molecule into the lipid bilayer of the spiroplasma membrane.
Wroblewski
H
H
Laboratoire de Microbiologie Fondamentale et Appliquée, Université de Rennes I, France.
Blanchard
A
A
Nyström
S
S
Wieslander
A
A
Thomas
D
D
D000215:Acylation; D000942:Antigens, Bacterial / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000302:isolation & purification*; D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D003902:Detergents / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008823:Micelles; D013045:Species Specificity; D013149:Spiroplasma / Q000032:analysis* / Q000276:immunology
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; D008081:Liposomes; D008823:Micelles; C028588:spiralin