CRYSTALLIZATION AND PRELIMINARY CRYSTALLOGRAPHIC DATA FOR ACETOHYDROXY ACID ISOMEROREDUCTASE FROM SPINACIA-OLERACEA

被引：10

作者：

DUMAS, R ^{[1
]}

JOB, D ^{[1
]}

DOUCE, R ^{[1
]}

PEBAYPEYROULA, E ^{[1
]}

COHENADDAD, C ^{[1
]}

机构：

[1] UNIV GRENOBLE 1, CEA, CNRS, INST BIOL STRUCT, F-38027 GRENOBLE 1, FRANCE

来源：

JOURNAL OF MOLECULAR BIOLOGY | 1994年 / 242卷 / 04期

关键词：

ACETOHYDROXY ACID ISOMEROREDUCTASE; CRYSTALLIZATION; PLANT; MAGNESIUM; HERBICIDE;

D O I：

10.1006/jmbi.1994.1602

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Acetohydroxy acid isomeroreductase (EC 1.1.1.86) is one of the enzymes involved in branched-chain amino acid biosynthesis. The enzyme from spinach (Spinacia oleracea) leaves has been crystallized using the hanging drop vapour diffusion method. The free enzyme crystallized from polyethylene glycol solutions, but these crystals were unsuitable for X-ray diffraction analysis. In the presence of NADPH, Mg2+ and a reaction intermediate analogue (2-dimethylphosphinoyl-2-hydroxy acetic acid (Hoe 704) or N-hydroxy-N-isopropyloxamate (IpOHA)), much better crystals were obtained. Crystals grown from ammonium sulphate belong to space group P2(1) with cell dimensions a = 193.78(7) Angstrom, b = 63.69(2) Angstrom, c = 112.84(1) Angstrom and beta = 121.22(1)degrees. The molecular mass of the protein, the volume of the unit cell, and crystal density measurements indicated that the asymmetric unit contains two dimers. X-ray diffraction patterns showed measurable reflections to beyond 2.5 Angstrom.

引用

页码：578 / 581

页数：4