GTP：α-D-甘露糖-1-磷酸鸟苷转移酶的纯化

Purification of GTP:alpha-D-mannose-1-phosphate guanyltransferase.

作者信息

Smoot J W, Serif G S

出版信息

Eur J Biochem. 1985 Apr 1;148(1):83-7. doi: 10.1111/j.1432-1033.1985.tb08810.x.

DOI:10.1111/j.1432-1033.1985.tb08810.x

Abstract

The enzyme GTP:alpha-D-mannose-1-phosphate guanylyltransferase from porcine thyroid tissue has been purified 69 900-fold on columns of blue-Sepharose, DEAE-Sepharose, phenyl-Sepharose and agarose-GTP affinity materials. Although it exhibits a tendency to aggregate, the enzyme travelled, upon sucrose velocity sedimentation, as a single oligomer with a molecular mass of 412 kDa. Michaelis constants were determined to be 1.0 microM, 1.0 mM, 3.5 microM and 0.4 microM for GDP-alpha-D-mannose, pyrophosphate, GTP and mannose-1-phosphate, respectively. The enzyme appears to be specific for the mannose moiety but will accept an inosine replacement for guanine and a deoxyribose replacement for ribose in GTP.

摘要

来自猪甲状腺组织的酶GTP：α-D-甘露糖-1-磷酸鸟苷酰转移酶已在蓝色琼脂糖凝胶、二乙氨基乙基琼脂糖凝胶、苯基琼脂糖凝胶和琼脂糖-GTP亲和材料柱上纯化了69900倍。尽管该酶有聚集的倾向，但在蔗糖速率沉降实验中，它以分子量为412 kDa的单一寡聚体形式移动。已确定GDP-α-D-甘露糖、焦磷酸、GTP和甘露糖-1-磷酸的米氏常数分别为1.0微摩尔、1.0毫摩尔、3.5微摩尔和0.4微摩尔。该酶似乎对甘露糖部分具有特异性，但在GTP中，它会接受肌苷取代鸟嘌呤以及脱氧核糖取代核糖。