Enrich C, Bachs O, Evans W H
Laboratory of Protein Structure, National Institute for Medical Research, London, U.K.
Biochem J. 1988 Nov 1;255(3):999-1005. doi: 10.1042/bj2550999.
The distribution of calmodulin-binding polypeptides in various rat liver subcellular fractions was investigated. Plasma-membrane, endosome, Golgi and lysosome fractions were prepared by established procedures. The calmodulin-binding polypeptides present in the subcellular fractions were identified by using an overlay technique after transfer from gels to nitrocellulose sheets. Distinctive populations of calmodulin-binding polypeptides were present in all the fractions examined except lysosomes. A major 115 kDa calmodulin-binding polypeptide of pI 4.3 was located to the endosome subfractions, and it emerges as a candidate endosome-specific protein. Partitioning of endosome fractions between aqueous and Triton X-114 phases indicated that the calmodulin-binding polypeptide was hydrophobic. Major calmodulin-binding polypeptides of 140 and 240 kDa and minor polypeptides of 40-60 kDa were present in plasma membranes. The distribution of calmodulin in the various endosome and plasma-membrane fractions was also analysed, and the results indicated that the amounts were high compared with those in the cytosol.
研究了钙调蛋白结合多肽在大鼠肝脏不同亚细胞组分中的分布情况。采用既定方法制备了质膜、内体、高尔基体和溶酶体组分。通过将凝胶中的蛋白质转移至硝酸纤维素膜上后使用覆盖技术,鉴定了亚细胞组分中存在的钙调蛋白结合多肽。除溶酶体外,在所检测的所有组分中均存在不同的钙调蛋白结合多肽群体。一种主要的115 kDa、pI为4.3的钙调蛋白结合多肽定位于内体亚组分,它成为一种候选的内体特异性蛋白。内体组分在水相和Triton X-114相之间的分配表明,该钙调蛋白结合多肽具有疏水性。质膜中存在140 kDa和240 kDa的主要钙调蛋白结合多肽以及40 - 60 kDa的次要多肽。还分析了钙调蛋白在各种内体和质膜组分中的分布,结果表明其含量与胞质溶胶中的相比很高。