通过CDR-琼脂糖亲和色谱法分离对Ca2+敏感和不敏感的腺苷酸环化酶以及钙依赖性调节蛋白（CDR）
Resolution of adenylate cyclase sensitive and insensitive to Ca2+ and calcium-dependent regulatory protein (CDR) by CDR-sepharose affinity chromatography.
作者信息
Westcott K R, La Porte D C, Storm D R
出版信息
Proc Natl Acad Sci U S A. 1979 Jan;76(1):204-8. doi: 10.1073/pnas.76.1.204.
Abstract
Partially purified adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] from bovine brain cortex was fractionated into two separate forms by calcium-dependent regulatory protein (CDR)-Sepharose affinity chromatography. The major form of the enzyme, comprising approximately 80% of the applied activity, did not bind to the affinity column in the presence of Ca2+ and was insensitive to the CDR. Approximately 20% of adenylate cyclase activity was absorbed to CDR-Sepharose in the presence of Ca2+. This activity was stimulated by Ca2+ and CDR. This study directly demonstrates that brain cortex contains Ca2+-CDR-sensitive and -insensitive forms of adenylate cyclase and indicates that CDR-Sepharose may be a useful tool for purification of adenylate cyclase. The Ca2+ -stimulated adenylate cyclase was purified at least 55-fold with a 13% yield.
摘要
通过钙依赖性调节蛋白（CDR）-琼脂糖亲和层析，将来自牛脑皮质的部分纯化的腺苷酸环化酶[ATP焦磷酸裂解酶（环化），EC 4.6.1.1]分离成两种不同的形式。该酶的主要形式约占所加活性的80%，在Ca2+存在下不与亲和柱结合，且对CDR不敏感。在Ca2+存在下，约20%的腺苷酸环化酶活性被CDR-琼脂糖吸附。该活性受到Ca2+和CDR的刺激。本研究直接证明脑皮质含有对Ca2+-CDR敏感和不敏感的腺苷酸环化酶形式，并表明CDR-琼脂糖可能是纯化腺苷酸环化酶的有用工具。受Ca2+刺激的腺苷酸环化酶纯化了至少55倍，产率为13%。
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Resolution of adenylate cyclase sensitive and insensitive to Ca2+ and calcium-dependent regulatory protein (CDR) by CDR-sepharose affinity chromatography.通过CDR-琼脂糖亲和色谱法分离对Ca2+敏感和不敏感的腺苷酸环化酶以及钙依赖性调节蛋白（CDR）
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