Coling D E, Salisbury J L
Center for NeuroSciences, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106.
J Protozool. 1992 May-Jun;39(3):385-91. doi: 10.1111/j.1550-7408.1992.tb01468.x.
Centrin is a major protein of the contractile striated flagellar roots of the green alga Tetraselmis striata. We present a newly modified procedure for the preparation of centrin in sufficient quantity and purity to allow for detailed biochemical characterization. We establish that centrin purified by differential solubility, followed by phenyl-Sepharose and DEAE-Sephacel chromatography is identical with the protein extracted directly from striated flagellar roots with regard to molecular weight, isoelectric point, and calcium-dependent behavior in SDS-PAGE. We also compare the biochemical properties of purified centrin with calmodulin isolated from Tetraselmis and calmodulin isolated from mammalian brain. Centrin can be fully distinguished from either algal or mammalian calmodulin on the basis of molecular weight, isoelectric point, calcium-dependent behavior in SDS-PAGE, proteolytic peptide maps, amino acid composition, ability to activate bovine brain phosphodiesterase, and reactivity with specific antibodies.
中心蛋白是绿藻条纹四鞭藻收缩横纹鞭毛根的一种主要蛋白质。我们提出了一种新的改良方法，用于制备足够数量和纯度的中心蛋白，以便进行详细的生化特性分析。我们确定，通过差异溶解度纯化，然后进行苯基琼脂糖和二乙氨基乙基琼脂糖凝胶层析纯化的中心蛋白，在分子量、等电点以及SDS-PAGE中的钙依赖性行为方面，与直接从横纹鞭毛根中提取的蛋白质相同。我们还比较了纯化的中心蛋白与从条纹四鞭藻中分离的钙调蛋白以及从哺乳动物大脑中分离的钙调蛋白的生化特性。基于分子量、等电点、SDS-PAGE中的钙依赖性行为、蛋白水解肽图谱、氨基酸组成、激活牛脑磷酸二酯酶的能力以及与特异性抗体的反应性，中心蛋白可以与藻类或哺乳动物的钙调蛋白完全区分开来。
