Tryptophan hydroxylase: Purification by affinity chromatography on calmodulin-Sepharose

被引：11

作者：

DSa, C

Arthur, R

Jennings, I

Cotton, RGH

Kuhn, DM

机构：

[1] WAYNE STATE UNIV,SCH MED,DEPT PSYCHIAT & BEHAV NEUROSCI,CELLULAR & CLIN NEUROBIOL PROGRAM,DETROIT,MI 48201

[2] ROYAL CHILDRENS HOSP,MURDOCH INST,OLIVE MILLER PROT LAB,PARKVILLE,VIC 3052,AUSTRALIA

[3] WAYNE STATE UNIV,SCH MED,CTR MOL MED & GENET,DETROIT,MI 48201

来源：

JOURNAL OF NEUROSCIENCE METHODS | 1996年 / 69卷 / 02期

关键词：

tryptophan hydroxylase; affinity chromatography; calmodulin; protein purification; brain;

D O I：

10.1016/S0165-0270(96)00034-9

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Tryptophan hydroxylase (EC 1.14.16.4; L-tryptophan, tetrahydropteridine:oxygen oxidoreductase (5-hydroxylating)) from rat mesencephalic tegmentum has been purified by sequential chromatography on Blue-Sepharose, DE-52, and calmodulin-Sepharose. The hydroxylase is excluded from Blue-Sepharose and is eluted from DE-52 with a step-wise NaCl gradient. Tryptophan hydroxylase binds to calmodulin-Sepharose in the presence of calcium and is eluted with either EGTA or calmodulin itself, but not with tryptophan. The purification scheme is rapid (5-6 h) and yields an enzyme with a specific activity of 225 nmol 5-HTP/mg min, representing a 400-fold purification with 7% recovery. The tryptophan hydroxylase preparation was judged to be > 95% pure using the present isolation procedure.

引用

页码：149 / 153

页数：5

共 28 条

[1] PURIFICATION AND CHARACTERIZATION OF THE BLUE GREEN RAT PHEOCHROMOCYTOMA-(PC12) TYROSINE-HYDROXYLASE WITH A DOPAMINE-FE(III) COMPLEX - REVERSAL OF THE ENDOGENOUS FEEDBACK INHIBITION BY PHOSPHORYLATION OF SERINE-40 [J].