TY - JOUR
T1 - Bordetella pertussis adenylate cyclase
T2 - Isolation and purification by calmodulin-sepharose 4B chromatography
AU - Friedman, R. L.
N1 - Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1987
Y1 - 1987
N2 - Purified preparations of adenylate cyclase were obtained from crude urea extracts of Bordetella pertussis by a one-step calmodulin affinity chromatography technique. Diluted extract was loaded onto the column and washed, and adenylate cyclase was eluted with 10mM EGTA [ethylene glycol-bis(β-aminoethyl ether)-N,N,N', N'-tetraacetic acid]. A 104-fold purification was accomplished in one step. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the affinity-purified adenylate cyclase was dissociated into one major protein band with an apparent molecular weight of 60,000 and a minor band at 200,000. The affinity-purified adenylate cyclase was observed (i) to have adenylate cyclase enzymatic activity which was activated by calmodulin, (ii) to bind 125I-calmodulin, and (iii) to be free of pertussis toxin as determined by in vivo and in vitro assays.
AB - Purified preparations of adenylate cyclase were obtained from crude urea extracts of Bordetella pertussis by a one-step calmodulin affinity chromatography technique. Diluted extract was loaded onto the column and washed, and adenylate cyclase was eluted with 10mM EGTA [ethylene glycol-bis(β-aminoethyl ether)-N,N,N', N'-tetraacetic acid]. A 104-fold purification was accomplished in one step. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the affinity-purified adenylate cyclase was dissociated into one major protein band with an apparent molecular weight of 60,000 and a minor band at 200,000. The affinity-purified adenylate cyclase was observed (i) to have adenylate cyclase enzymatic activity which was activated by calmodulin, (ii) to bind 125I-calmodulin, and (iii) to be free of pertussis toxin as determined by in vivo and in vitro assays.
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U2 - 10.1128/iai.55.1.129-134.1987
DO - 10.1128/iai.55.1.129-134.1987
M3 - Article
C2 - 2878883
AN - SCOPUS:0023074159
VL - 55
SP - 129
EP - 134
JO - Infection and Immunity
JF - Infection and Immunity
SN - 0019-9567
IS - 1
ER -