Effects of calcium on recombinant bovine chromogranin A.
Effects of calcium on recombinant bovine chromogranin A.
复制标题
钙对重组牛嗜铬粒蛋白A的影响。
DOI：
10.1002/pro.5560011208
复制
发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Nieves,E
中科院分区：
文献类型：
--
作者：
Angeletti,RH;Ali,G;Shen,N;Gee,P;Nieves,E
Bovine chromogranin A, the acidic calcium‐binding protein characteristic of endocrine secretory vesicles, has been expressed inEscherichia coliusing the pET3a vector system under T7 polymerase control. The expressed protein is located in the bacterial cytosol and can be purified from bacterial proteins by a heat treatment step, followed by gel filtration, anion‐exchange, and reversed‐phase chromatography.纯化的重组嗜铬粒蛋白 A 的表观 MT 为 ca。 72,000 by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, in spite of its 432‐amino acid polypeptide chain, consistent with observations on natural chromogranin A. The primary structure has been confirmed by mass spectral analysis of tryptic peptides, by Edman degradation of the intact protein, and by immunoreactivity with sequence‐specific antibodies.圆二色光谱分析显示了 pH 值和浓度依赖性光谱。光谱在 5 至 40 μM 范围内依赖于 Ca2+。
Bovine chromogranin A, the acidic calcium‐binding protein characteristic of endocrine secretory vesicles, has been expressed inEscherichia coliusing the pET3a vector system under T7 polymerase control. The expressed protein is located in the bacterial cytosol and can be purified from bacterial proteins by a heat treatment step, followed by gel filtration, anion‐exchange, and reversed‐phase chromatography. The purified recombinant chromogranin A has an apparentMTof ca. 72,000 by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, in spite of its 432‐amino acid polypeptide chain, consistent with observations on natural chromogranin A. The primary structure has been confirmed by mass spectral analysis of tryptic peptides, by Edman degradation of the intact protein, and by immunoreactivity with sequence‐specific antibodies. Analysis by circular dichroism spectroscopy shows pH‐ and concentration‐dependent spectra. The spectra are Ca2+‐dependent from 5 to 40 μM.