A Cl(-)-translocating adenosinetriphosphatase in Acetabularia acetabulum. 1. Purification and characterization of a novel type of adenosinetriphosphatase that differs from chloroplast F1 adenosinetriphosphatase.
A Cl(-)-translocating adenosinetriphosphatase in Acetabularia acetabulum. 1. Purification and characterization of a novel type of adenosinetriphosphatase that differs from chloroplast F1 adenosinetriphosphatase.
复制标题
髋臼中的 Cl(-)-转位腺苷三磷酸酶。
作者：
M. Ikeda;R. Schmid;D. Oesterhelt
用壬酰- n-甲基葡萄糖酰胺从髋臼膜中溶解atp酶，并用离子交换和凝胶渗透色谱法纯化。分离出三种ATPase， Mono Q-I， -II和-III。Mono q - 1组分的活性不稳定，不能准确测定。Mono Q-II和-III组分的比活性分别为0.6和6单位/mg蛋白质。通过sds -聚丙烯酰胺凝胶电泳、等电聚焦和多肽图谱分析，发现Mono Q-II和-III组分由相同的多肽组成，分子量分别为54K （a亚基）和50K （b亚基）。组分Mono Q-II和-III具有以下催化性能：pH为6.0时最优；底物特异性，ATP = GTP = ITP远大于UTP = CTP (Km为ATP 0.6 mM)；二价阳离子要求，Mn2+ = Mg2+大于Co2+大于Zn2+，远远大于Ca2+、Ni2+。这两种活性均被一价阴离子抑制，而一价阳离子既无抑制作用，也无刺激作用。正钒酸盐在1mm时将两者的活性抑制到50%，叠氮化物在100微米时的抑制率为95%。Mono Q-III组分与[γ - 32p]ATP孵育后形成酶-磷酸复合物。cf1 - atp酶亚复合物从同一生物体中分离出来，并与Mono Q-III组分进行比较。数据支持Mono Q-III分数与CF1-ATPase的差异。
ATPases were solubilized from membranes of Acetabularia acetabulum using nonanoyl-N-methylgluconamide and purified by ion-exchange and gel permeation chromatography. Three fractions of ATPase, Mono Q-I, -II, and -III, were separated. Activity in fraction Mono Q-I was very labile and could not be accurately determined. Fractions Mono Q-II and -III had specific activities of 0.6 and 6 units/mg of protein, respectively. By SDS-polyacrylamide gel electrophoresis, isoelectric focusing, and peptide mapping, it was shown that fractions Mono Q-II and -III consisted of the same polypeptides with molecular masses of 54K (a-subunit) and 50K (b-subunit). Fractions Mono Q-II and -III had the following catalytic properties: pH optimum at 6.0; substrate specificity, ATP = GTP = ITP much greater than UTP = CTP (Km for ATP 0.6 mM); divalent cation requirement, Mn2+ = Mg2+ greater than Co2+ greater than Zn2+ much greater than Ca2+, Ni2+. Both activities were inhibited by monovalent anions, while monovalent cations had neither inhibitory nor stimulatory effects. Orthovanadate inhibited both activities to 50% at 1 mM, and the most effective inhibitor of both was azide (95% inhibition at 100 microM). An enzyme-phosphate complex was formed after incubation of fraction Mono Q-III with [gamma-32P]ATP. The CF1-ATPase subcomplexes were isolated from the same organism and compared with the fraction Mono Q-III. Data supported the difference of fraction Mono Q-III from CF1-ATPase.
一种在十二烷基硫酸钠-聚丙烯酰胺凝胶中使用 N-氯代琥珀酰亚胺/尿素和肽银染色进行部分肽图谱分析的新方法。
影响因子：
2.9
作者：
Lischwe,MA;Ochs,D
通讯作者：
Ochs,D