BORDETELLA-PERTUSSIS ADENYLATE-CYCLASE - ISOLATION AND PURIFICATION BY CALMODULIN-SEPHAROSE-4B CHROMATOGRAPHY
BORDETELLA-PERTUSSIS ADENYLATE-CYCLASE - ISOLATION AND PURIFICATION BY CALMODULIN-SEPHAROSE-4B CHROMATOGRAPHY
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DOI：
10.1128/iai.55.1.129-134.1987
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发表时间：
1987-01-01
影响因子：
3.1
通讯作者：
FRIEDMAN, RL
中科院分区：
文献类型：
--
作者：
FRIEDMAN, RL
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(.beta.-aminoethyl)-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 adenylase 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.
