产朊假丝酵母NAD⁺激酶：纯化、性质及亲和凝胶研究

Candida utilis NAD+ kinase: purification, properties and affinity gel studies.

Butler J R, McGuinness E T

Int J Biochem. 1982;14(9):839-44. doi: 10.1016/0020-711x(82)90106-9.

Abstract

NAD+ kinase (ATP:NAD+ 2' phosphotransferase, EC 2.7.1.23) has been purified to apparent enzymic homogeneity on Blue Sepharose CL-6B. 2. The molecular weight of the active species is about 260,000 as determined by PAGE and gel chromatography. Protein staining (PAGE) revealed minor bands with molecular weight values of 40,000, 140,000 and 550,000. Subunit studies (SDS-PAGE) gave evidence of a single band of molecular weight approximately 32,000. 3. On the basis of the release patterns of this enzyme from several affinity gels, an elution diagram is proposed as a device to assess the contribution of any of the several displacing agents that can be used to manipulate the desorption of a (enzyme) ligate from an immobilized ligand.

摘要

NAD⁺激酶（ATP：NAD⁺ 2'-磷酸转移酶，EC 2.7.1.23）已通过Blue Sepharose CL-6B纯化至表观酶学均一性。2. 通过PAGE和凝胶色谱法测定，活性物质的分子量约为260,000。蛋白质染色（PAGE）显示分子量值为40,000、140,000和550,000的次要条带。亚基研究（SDS-PAGE）证明有一条分子量约为32,000的单一条带。3. 根据该酶从几种亲和凝胶上的释放模式，提出了一种洗脱图，作为一种评估任何可用于操纵固定化配体上（酶）连接物解吸的几种置换剂贡献的工具。