TY - JOUR
T1 - Isolation of high-specific-activity subunits of cholera toxin by reversed-phase high-performance liquid chromatography
AU - Pearson, Stephen D.
AU - Dixon, James D.
AU - Northwehr, Steven F.
AU - Kurosky, Alexander
N1 - Funding Information:
This investigation was supported by National Institutes of Health Research Grants GM 29039 and CA 17701. S. D. Pearson is a James W. McLaughlin Predoctoral Fellow. We wish to thank Horace Kelso for expert technical assistance. Discussions with Randall M. Goldblum were much appreciated.
PY - 1986
Y1 - 1986
N2 - 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 α-, γ-, and β-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 α-chain, or carboxymethyl α-chain was also determined to be reasonably high compared to that of cholera toxin or commercially prepared A subunit.
AB - 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 α-, γ-, and β-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 α-chain, or carboxymethyl α-chain was also determined to be reasonably high compared to that of cholera toxin or commercially prepared A subunit.
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U2 - 10.1016/0021-9673(86)80095-4
DO - 10.1016/0021-9673(86)80095-4
M3 - Article
C2 - 3016001
AN - SCOPUS:0022556355
SN - 0021-9673
VL - 359
SP - 413
EP - 421
JO - Journal of Chromatography A
JF - Journal of Chromatography A
IS - C
ER -