Isolation of high-specific-activity subunits of cholera toxin by reversed-phase high-performance liquid chromatography.
Facile, rapid procedures for the separation of active cholera toxin subunits were developed, based on high-performance liquid chromatography (HPLC) with a Nucleosil C8 reversed-phase column. These procedures were capable of completely resolving subunits A and B as well as S-carboxymethylated or reduced alpha-, gamma-, and beta-chains. The binding of HPLC-purified B subunit to GM1 ganglioside was essentially identical to that of cholera toxin when compared on a molar basis. The adenosine 5'-diphosphate-ribosyltransferase activity of HPLC-purified A subunit, reduced alpha-chain, or carboxymethyl alpha-chain was also determined to be reasonably high compared to that of cholera toxin or commercially prepared A subunit.
D000478:Alkylation; D000596:Amino Acids / Q000032:analysis; D002772:Cholera Toxin / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005677:G(M1) Ganglioside / Q000032:analysis; D009713:Nucleotidyltransferases / Q000032:analysis* / Q000037:antagonists & inhibitors; D010084:Oxidation-Reduction; D011065:Poly(ADP-ribose) Polymerases
D000596:Amino Acids; D005677:G(M1) Ganglioside; D002772:Cholera Toxin; D011065:Poly(ADP-ribose) Polymerases; D009713:Nucleotidyltransferases