Expression and purification of a mutant human growth hormone that is resistant to proteolytic cleavage by thrombin, plasmin and human plasma in vitro

被引：5

作者：

Alam, KSM ^{[1
]}

Morimoto, M ^{[1
]}

Yoshizato, H ^{[1
]}

Fujikawa, T ^{[1
]}

Furukawa, K ^{[1
]}

Tanaka, M ^{[1
]}

Nakashima, K ^{[1
]}

机构：

[1] Mie Univ, Fac Med, Dept Biochem, Tsu, Mie 514, Japan

来源：

JOURNAL OF BIOTECHNOLOGY | 1998年 / 65卷 / 2-3期

关键词：

human growth hormone; mutation; proteolytic insensitivity; plasmin; thrombin; human plasma;

D O I：

10.1016/S0168-1656(98)00113-8

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The region having a sequence from amino acid 134 to 150 in human growth hormone (hGH) is known to be cleaved by proteases in human plasma, plasmin and thrombin. In this study, oligonucleotide primer-directed mutagenesis was used to produce recombinant mutant hGHs resistant to limited proteolysis by these proteases. Substitution of Arg(134) and Thr(135) of hGH with Asp(134) and Pro(135) yielded a thrombin-resistant hGH mutant: and substitution of Arg(134), Thr(135) and Lys(140) With Asp(134), Pro(135) and Ala(140) yielded a plasmin-resistant hGH mutant. The latter mutant hGH was also insensitive to in vitro proteolysis by human plasma incubated for 7 days. These alterations in amino acid residues of hGH did not disrupt its biological conformation and retained full growth promoting activities on rat Nb2 cells and human T-47D breast cancer cells. (C) 1998 Elsevier Science B.V. All rights reserved.

引用

页码：183 / 190

页数：8

共 23 条

[1] TEMPERATURE-SENSITIVE MUTATIONS OF BACTERIOPHAGE-T4 LYSOZYME OCCUR AT SITES WITH LOW MOBILITY AND LOW SOLVENT ACCESSIBILITY IN THE FOLDED PROTEIN [J].