SITE-DIRECTED MUTAGENESIS IN THE ACTIVE-SITE OF THE HERPES-SIMPLEX VIRUS TYPE-1 THYMIDINE KINASE GENE

被引：8

作者：

FETZER, J ^{[1
]}

FOLKERS, G ^{[1
]}

MULLER, I ^{[1
]}

KEIL, GM ^{[1
]}

机构：

[1] FED RES CTR VIRUS DIS ANIM, TUBINGEN, GERMANY

来源：

VIRUS GENES | 1993年 / 7卷 / 02期

关键词：

HSV-1 THYMIDINE KINASE; SITE-DIRECTED MUTAGENESIS; HSV-1 TK ANTISERUM;

D O I：

10.1007/BF01702400

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The thymidine kinase (TK) of herpes simplex virus type 1 (HSV-1) contains three regions of homology to other ATP utilizing enzymes. We have altered one region of the protein, which seems to play an important role in phosphorylating substrates by site-directed mutagenesis. When the aspartate 162 was changed to asparagine, the enzyme lost its activity. To identify the inactive protein, expressed by a vaccinia vector in eukaryotic cells, a monospecific antiserum against a bacterial tryptophan E-HSV-1 TK fusion protein was made. These results support the suggestion that aspartate 162 is essential for the enzymatic activity.

引用

收藏

页码：205 / 209

页数：5

相关论文

共 50 条

[21] Potential active-site residues in polyneuridine aldehyde esterase, a central enzyme of indole alkaloid biosynthesis, by modelling and site-directed mutagenesis [J].

Mattern-Dogru, E ;

Ma, XY ;

Hartmann, J ;

Decker, H ;

Stöckigt, J .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 2002, 269 (12) :2889-2896

[22] Significance of amino acid substitutions in the thymidine kinase gene of herpes simplex virus type 1 for resistance [J].

Sauerbrei, Andreas ;

Liermann, Kristin ;

Bohn, Kathrin ;

Henke, Andreas ;

Zell, Roland ;

Gronowitz, Simon ;

Wutzler, Peter .

ANTIVIRAL RESEARCH, 2012, 96 (02) :105-107

[23] IDENTIFICATION OF ACTIVE-SITE LYSYL RESIDUES OF PHENYLALANINE DEHYDROGENASE BY CHEMICAL MODIFICATION WITH METHYL ACETYL PHOSPHATE COMBINED WITH SITE-DIRECTED MUTAGENESIS [J].

KATAOKA, K ;

TANIZAWA, K ;

FUKUI, T ;

UENO, H ;

YOSHIMURA, T ;

ESAKI, N ;

SODA, K .

JOURNAL OF BIOCHEMISTRY, 1994, 116 (06) :1370-1376

[24] Site-directed mutagenesis of the active site of diacylglycerol kinase α:: calcium and phosphatidylserine stimulate enzyme activity via distinct mechanisms [J].

Abe, T ;

Lu, XL ;

Jiang, Y ;

Boccone, CE ;

Qian, SM ;

Vattem, KM ;

Wek, RC ;

Walsh, JP .

BIOCHEMICAL JOURNAL, 2003, 375 :673-680

[25] Role of the amino acid invariants in the active site of MurG as evaluated by site-directed mutagenesis [J].

Crouvoisier, Muriel ;

Auger, Genevieve ;

Blanot, Didier ;

Mengin-Lecreulx, Dominique .

BIOCHIMIE, 2007, 89 (12) :1498-1508

[26] Molecular modelling and site-directed mutagenesis of the active site of endothelin-converting enzyme [J].

Sansom, CE ;

Hoang, MV ;

Turner, AJ .

PROTEIN ENGINEERING, 1998, 11 (12) :1235-1241

[27] Valine dehydrogenase from Streptomyces albus:: gene cloning, heterologous expression and identification of active site by site-directed mutagenesis [J].

Hyun, CG ;

Kim, SS ;

Park, KH ;

Suh, JW .

FEMS MICROBIOLOGY LETTERS, 2000, 182 (01) :29-34

[28] Site-directed mutagenesis studies of the aromatic residues at the active site of a lipase from Malassezia globosa [J].

Gao, Chongliang ;

Lan, Dongming ;

Liu, Lu ;

Zhang, Houjin ;

Yang, Bo ;

Wang, Yonghua .

BIOCHIMIE, 2014, 102 :29-36

[29] SITE-DIRECTED MUTAGENESIS OF LA PROTEASE - A CATALYTICALLY ACTIVE SERINE RESIDUE [J].

AMERIK, AY ;

ANTONOV, VK ;

GORBALENYA, AE ;

KOTOVA, SA ;

ROTANOVA, TV ;

SHIMBAREVICH, EV .

FEBS LETTERS, 1991, 287 (1-2) :211-214

[30] Substitution of conserved glycine with threonine in the active site of OsTrx23 by site-directed mutagenesis [J].

Nashta, Mitra Roodgar ;

Shahpiri, Azar .

MINERVA BIOTECNOLOGICA, 2016, 28 (04) :219-223

← 1 2 3 4 5 →