Expression of human tyrosine hydroxylase type I in Escherichia coli as a protease-cleavable fusion protein

被引：10

作者：

Nakashima, A

Mori, K

Nagatsu, T

Ota, A ^{[1
]}

机构：

[1] Fujita Hlth Univ, Sch Med, Dept Physiol, Aichi 4701192, Japan

[2] Fujita Hlth Univ, Inst Comprehens Med Sci, Div Mol Genet, Aichi 4701192, Japan

来源：

JOURNAL OF NEURAL TRANSMISSION | 1999年 / 106卷 / 9-10期

关键词：

human tyrosine hydroxylase type 1; N-terminal amino acid-deleted mutant; maltose-binding protein fusion;

D O I：

10.1007/s007020050202

中图分类号：

R74 [神经病学与精神病学];

学科分类号：

摘要：

Wild-type and N-terminal 35-, 38-, and 44-amino acid-deleted mutants of human tyrosine hydroxylase type 1 (hTH1) fused to maltose-binding protein via the target sequence for a restriction protease were expressed in Escherichia coli and purified. The fused protein was treated with the restriction protease factor Xa or enterokinase to isolate hTH1 from the fused form. The treatment of fused wild-type and 35-amino acid-deleted mutant with factor Xa and enterokinase caused non-specific cleavages in the vicinity of the phosphorylation sites, Ser(19) and Ser(40), due to the flexible conformation of the N-terminus of hTH1.

引用

页码：819 / 824

页数：6

共 14 条

[1]

ABATE C, 1991, J MOL NEUROSCI, V2, P203

[2] Phosphorylation of recombinant human phenylalanine hydroxylase: Effect on catalytic activity, substrate activation and protection against non-specific cleavage of the fusion protein by restriction protease [J].