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] IDENTIFICATION OF ACTIVE-SITE LYSYL RESIDUES OF PHENYLALANINE DEHYDROGENASE BY CHEMICAL MODIFICATION WITH METHYL ACETYL PHOSPHATE COMBINED WITH SITE-DIRECTED MUTAGENESIS
    KATAOKA, K
    TANIZAWA, K
    FUKUI, T
    UENO, H
    YOSHIMURA, T
    ESAKI, N
    SODA, K
    JOURNAL OF BIOCHEMISTRY, 1994, 116 (06) : 1370 - 1376
  • [22] Significance of amino acid substitutions in the thymidine kinase gene of herpes simplex virus type 1 for resistance
    Sauerbrei, Andreas
    Liermann, Kristin
    Bohn, Kathrin
    Henke, Andreas
    Zell, Roland
    Gronowitz, Simon
    Wutzler, Peter
    ANTIVIRAL RESEARCH, 2012, 96 (02) : 105 - 107
  • [23] Potential active-site residues in polyneuridine aldehyde esterase, a central enzyme of indole alkaloid biosynthesis, by modelling and site-directed mutagenesis
    Mattern-Dogru, E
    Ma, XY
    Hartmann, J
    Decker, H
    Stöckigt, J
    EUROPEAN JOURNAL OF BIOCHEMISTRY, 2002, 269 (12): : 2889 - 2896
  • [24] Role of the amino acid invariants in the active site of MurG as evaluated by site-directed mutagenesis
    Crouvoisier, Muriel
    Auger, Genevieve
    Blanot, Didier
    Mengin-Lecreulx, Dominique
    BIOCHIMIE, 2007, 89 (12) : 1498 - 1508
  • [25] Site-directed mutagenesis of the active site of diacylglycerol kinase α:: calcium and phosphatidylserine stimulate enzyme activity via distinct mechanisms
    Abe, T
    Lu, XL
    Jiang, Y
    Boccone, CE
    Qian, SM
    Vattem, KM
    Wek, RC
    Walsh, JP
    BIOCHEMICAL JOURNAL, 2003, 375 : 673 - 680
  • [26] Molecular modelling and site-directed mutagenesis of the active site of endothelin-converting enzyme
    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
    Hyun, CG
    Kim, SS
    Park, KH
    Suh, JW
    FEMS MICROBIOLOGY LETTERS, 2000, 182 (01) : 29 - 34
  • [28] Site-directed mutagenesis of the essential arginine of the formate dehydrogenase active centre
    Galkin, AG
    Kutsenko, AS
    Bajulina, NP
    Esipova, NG
    Lamzin, VS
    Mesentsev, AV
    Shelukho, DV
    Tikhonova, TV
    Tishkov, VI
    Ustinnikova, TB
    Popov, VO
    BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 2002, 1594 (01): : 136 - 149
  • [29] Site-directed mutagenesis of key amino acids in the active site of amylosucrase from Neisseria polysaccharea
    Albenne, C
    De Montalk, GP
    Monsan, P
    Skov, L
    Mirza, O
    Gajhede, M
    Remaud-Simeon, M
    BIOLOGIA, 2002, 57 : 119 - 128
  • [30] SITE-DIRECTED MUTAGENESIS OF LA PROTEASE - A CATALYTICALLY ACTIVE SERINE RESIDUE
    AMERIK, AY
    ANTONOV, VK
    GORBALENYA, AE
    KOTOVA, SA
    ROTANOVA, TV
    SHIMBAREVICH, EV
    FEBS LETTERS, 1991, 287 (1-2) : 211 - 214
12345