A specific soluble protein from the catecholamine storage vesicles of bovine adrenal medulla. I. Purification and chemical characterization.
A specific soluble protein from the catecholamine storage vesicles of bovine adrenal medulla. I. Purification and chemical characterization.
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来自牛肾上腺髓质儿茶酚胺储存囊泡的特定可溶性蛋白质。
DOI：
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发表时间：
1967
影响因子：
3.6
通讯作者：
Norman Kirshner
中科院分区：
文献类型：
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作者：
W. Smith;Norman Kirshner
用deae -纤维素和Sephadex G-200层析纯化了时间型儿茶酚胺储存囊泡的可溶性蛋白。主要的蛋白质部分（S1）似乎是高度纯化的：它在超离心机中以单一的均匀部分沉积；在不同的pH值下，它在电泳上除了少数例外地给出了一个单一的蛋白质点，并且它的氨基酸组成和胰蛋白酶消化的指纹图谱在不同的制备过程中是恒定的。免疫和酶学分析表明，该蛋白含有不到1%的高免疫原性污染物，其中一种是多巴胺-β-氧化酶。在Sephadex G-200色谱柱上，除主要蛋白组分外，还获得了另外两个蛋白峰。其中一种（S2）在超离心中表现均匀，在蛋白质浓度为3 mg/ml时，其S25，w为1.4。这个分数的纯度低于S1。氨基酸组成、胰蛋白酶消化指纹图谱和S1分子量的研究表明，该蛋白可能由两个相同的亚基组成，分子量为40000。时间数据符合(a)单肽链具有一个链内二硫键，或(b)两个或多个不同链具有一个链内或链间二硫键的单位结构。对S1的时间结合能力的研究表明，它本身或在Mg++和ATP存在的情况下，不能结合足够数量的儿茶酚胺，从而不能解释储存囊泡内相当一部分儿茶酚胺的稳定性。
The soluble proteins obtained by osmotic lysis of time catecholamine storage vesicles have been purified by chromatography on DEAE-cellulose and Sephadex G-200. The major protein fraction (S1) appears to be highly purified: it sedimented as a single homogeneous fraction in the ultracentrifuge; it gave, with few exceptions, a single protein spot upon electrophoresis at different pH values, and its amino acid composition and fingerprints of tryptic digests were constant from preparation to preparation. Immunological and enzymic analysis of this protein indicated that it contained less than 1% of highly immunogenic contaminants, one of which was dopamine-β-oxidase. In addition to the major protein fraction, two other protein peaks were obtained from the Sephadex G-200 column. One of these (S2) appeared homogeneous in the ultracentrifuge and had an S25,w of 1.4 at a protein concentration of 3 mg/ml. This fraction was less pure than S1. Studies of the amino acid composition, fingerprints of tryptic digests and the molecular weight of S1 indicate that the protein may consist of two identical subunits having a molecular weight of 40,000. Time data are consistent with a unit structure of (a) a single peptide chain with one intrachain disulfide bond, or (b) two or more different chains with one intra- or one interchain disulfide bond. Studies of time binding capacity of S1 showed that it did not, by itself, or in the presence of Mg++ and ATP, bind sufficient amounts of catecholamines to be able to account for the stability of a significant fraction of the catecholamines within the storage vesicles.