RAPID PURIFICATION AND CHARACTERIZATION OF HIV-1 REVERSE-TRANSCRIPTASE AND RNASEH ENGINEERED TO INCORPORATE A C-TERMINAL TRIPEPTIDE ALPHA-TUBULIN EPITOPE

被引：49

作者：

STAMMERS, DK

TISDALE, M

COURT, S

PARMAR, V

BRADLEY, C

ROSS, CK

机构：

[1] Department of Molecular Sciences, The Wellcome Research Laboratories, Beckenham, Kent BR3 3BS, Longley Court

来源：

FEBS LETTERS | 1991年 / 283卷 / 02期

关键词：

HIV REVERSE TRANSCRIPTASE; RNASEH; PROTEIN ENGINEERING; ALPHA-TUBULIN EPITOPE; IMMUNOAFFINITY PURIFICATION;

D O I：

10.1016/0014-5793(91)80613-8

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The C-termini of p66 and p51 forms of HIV-1 reverse transcriptase have been engineered to contain a Glu-Glu-Phe sequence recognized by a monoclonal antibody to alpha-tubulin, YL1/2. Mutated RTs were purified in a single step using peptide elution from columns of immobilized YL1/2. The known sequence requirements of the YL1/2 epitope are consistent with protein eluting from the column with an intact C-terminus. Kinetic parameters of these mutated RTs are essentially unchanged from wild-type enzyme. The p15 RNaseH domain has been purified using this method and shown to have low enzyme activity compared to the parental p66 subunit.

引用

页码：298 / 302

页数：5

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