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来自酿酒酵母的胱硫醚β-合酶催化核心的纯化、结晶及初步晶体学分析。

Purification, crystallization and preliminary crystallographic analysis of the catalytic core of cystathionine β-synthase from Saccharomyces cerevisiae.

作者信息

Ereño-Orbea June, Majtan Tomas, Oyenarte Iker, Kraus Jan P, Martínez-Cruz Luis Alfonso

机构信息

Structural Biology Unit, CIC bioGUNE, Parque Tecnológico de Bizkaia, Edificio 800, 48160 Derio, Bizkaia , Spain.

Department of Pediatrics, University of Colorado School of Medicine, Aurora, CO 80045, USA.

出版信息

Acta Crystallogr F Struct Biol Commun. 2014 Mar;70(Pt 3):320-5. doi: 10.1107/S2053230X14001502. Epub 2014 Feb 19.

Abstract

Cystathionine β-synthase (CBS; EC 4.2.1.22) catalyzes the condensation of homocysteine and serine to form cystathionine, with the release of water. In humans, deficiency in CBS activity is the most common cause of hyperhomocysteinaemia and homocystinuria. More than 160 pathogenic mutations in the human CBS gene have been described to date. Here, the purification and preliminary crystallographic analysis of the catalytic core of CBS from Saccharomyces cerevisiae (ScCBS) is described which, in contrast to other eukaryotic CBSs, lacks the N-terminal haem-binding domain and is considered to be a useful model for investigation of the pyridoxal-5'-phosphate-mediated reactions of human CBS (hCBS). The purified protein yielded two different crystal forms belonging to space groups P41212 and P212121, with unit-cell parameters a = b = 72.390, c = 386.794 Å and a = 58.156, b = 89.988, c = 121.687 Å, respectively. Diffraction data were collected to 2.7 and 3.1 Å resolution, respectively, using synchrotron radiation. Preliminary analysis of the X-ray data suggests the presence of ScCBS homodimers in both types of crystals.

摘要

胱硫醚β-合酶(CBS;EC 4.2.1.22)催化同型半胱氨酸和丝氨酸缩合形成胱硫醚,并释放出水。在人类中,CBS活性缺乏是高同型半胱氨酸血症和同型胱氨酸尿症最常见的病因。迄今为止,已描述了人类CBS基因中的160多种致病突变。本文描述了酿酒酵母CBS(ScCBS)催化核心的纯化及初步晶体学分析,与其他真核生物CBS不同,ScCBS缺乏N端血红素结合结构域,被认为是研究人类CBS(hCBS)吡哆醛-5'-磷酸介导反应的有用模型。纯化后的蛋白质产生了两种不同的晶体形式,分别属于空间群P41212和P212121,晶胞参数分别为a = b = 72.390,c = 386.794 Å和a = 58.156,b = 89.988,c = 121.687 Å。使用同步辐射分别收集了分辨率为2.7和3.1 Å的衍射数据。X射线数据的初步分析表明,两种晶体中均存在ScCBS同型二聚体。

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