Bordetella pertussis adenylate cyclase. Purification, characterization, and radioimmunoassay.
Bordetella pertussis adenylate cyclase. Purification, characterization, and radioimmunoassay.
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DOI：
10.1016/s0021-9258(18)66710-9
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发表时间：
1986-12
期刊：
影响因子：
--
通讯作者：
Daniel Ladant;Colette Brezins;Jean-Michel Alonsos;Isabelle CrenonS;Nicole Guiso
中科院分区：
文献类型：
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作者：
Daniel Ladant;Colette Brezins;Jean-Michel Alonsos;Isabelle CrenonS;Nicole Guiso
细胞外腺苷酸环化酶纯化为游离酶或与钙调蛋白复合物。纯化后的酶具有1600 μ mol的cAMP min-1 X mg-1的比活性，存在于45和43 kDa的两种分子形式下，它们在结构上明显相关。钙调素显著增加腺苷酸环化酶对胰蛋白酶失活的抗性。尽管胰蛋白酶裂解了腺苷酸环化酶-钙调蛋白复合物，但被消化的片段仍以非共价相互作用的方式保持活性构象。特异性小鼠抗腺苷酸环化酶抗体抑制腺苷酸环化酶活性，并用于开发特异性放射免疫测定，可在培养上清中检测到5ng的腺苷酸环化酶。
The extracellular adenylate cyclase of Bordetella pertussis was purified either as a free enzyme or as a complex with calmodulin. The purified enzyme has a specific activity of 1600 mumol of cAMP min-1 X mg-1 and exists under two molecular forms of 45 and 43 kDa which are apparently structurally related. Calmodulin increased considerably the resistance of adenylate cyclase to inactivation by trypsin. Although trypsin cleaved the adenylate cyclase-calmodulin complex, the digested fragments remained associated by noncovalent interactions in an active conformation. Specific mouse anti-adenylate cyclase antibodies inhibit adenylate cyclase activity and were used to develop a specific radioimmunoassay that allows detection of as little as 5 ng of adenylate cyclase in culture supernatants.