Cell surface proteins from Shigella dysenteriae type 1.
A simple extraction procedure was used for preparing cell surface proteins (CSPs) from Shigella dysenteriae type 1. The preparations obtained using either buffer or water extractions were free from lipopolysaccharide (LPS), as well as cytoplasmic and periplasmic proteins. By SDS-PAGE, about 25 polypeptides were detected, and Western-blot analysis recognised 15 polypeptide antigens. When analysed by crossed immunoelectrophoresis, using anti-Shigella dysenteriae type 1 rabbit sera, 18 antigenic bands were identified. Proteins obtained by this method were found to be highly immunogenic in rabbits. The cell-surface proteins were compared to outer membrane proteins (OMPs) obtained from the S. dysenteriae type 1 strain by a standard procedure involving lysozyme-EDTA extraction, sucrose density centrifugation, and detergent treatment. They were found to contain periplasmic, cytoplasmic, and lipopolysaccharide contaminants. Thus, the procedure described here offers a quick and simple alternative for obtaining relatively pure cell surface proteins from Shigella dysenteriae type 1. This method will be useful when immunogenically active proteins free from other cellular components are required for studies.
Qadri
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F
International Centre for Diarrhoeal Disease Research Bangladesh, Dhaka.
Raqib
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R
Husain
I A
IA
Ciznár
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I
D000942:Antigens, Bacterial / Q000032:analysis / Q000302:isolation & purification; D000954:Antigens, Surface / Q000032:analysis / Q000302:isolation & purification; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000032:analysis* / Q000302:isolation & purification; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional; D008070:Lipopolysaccharides / Q000032:analysis / Q000302:isolation & purification; D008565:Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification; D012762:Shigella dysenteriae / Q000032:analysis* / Q000276:immunology / Q000648:ultrastructure
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D008565:Membrane Proteins