AFFINITY MODIFICATION OF HUMAN-IMMUNODEFICIENCY-VIRUS REVERSE-TRANSCRIPTASE AND DNA-TEMPLATE BY PHOTOREACTIVE DCTP ANALOGS

被引：29

作者：

DORONIN, SV ^{[1
]}

DOBRIKOV, MI ^{[1
]}

BUCKLE, M ^{[1
]}

ROUX, P ^{[1
]}

BUC, H ^{[1
]}

LAVRIK, OI ^{[1
]}

机构：

[1] RUSSIAN ACAD SCI,INST BIOORGAN CHEM,NOVOSIBIRSK 630090,RUSSIA

来源：

FEBS LETTERS | 1994年 / 354卷 / 02期

关键词：

HIV REVERSE TRANSCRIPTASE; DCTP ANALOG; PHOTOAFFINITY LABELING;

D O I：

10.1016/0014-5793(94)01110-9

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：

New base-substituted analogs of dCTP containing an azido group have been synthesized and applied to a selective photoaffinity modification of HIV-RT (p66/p51 heterodimer). The labeling of only the 66 kDa subunit of HIV-RT was detected when the enzyme was first irradiated with the analogs and then template (5'-(d)GGTTAAATAAAATAGTAAGAATGTATAGCCCCTACCA-3') and 5' P-32 end-labeled 3'-(d)TTACATATCGGGGATGGT-5' primer were added. The 5' P-32 end-labeled primer elongated by dCTP analogs in the presence of both HIV-RT and DNA template is able to modify both subunits of HIV-RT and DNA template. This way of specific cross-linking to both DNA (RNA) template and HIV-RT opens up new possibilities to study the HIV-RT active site.

引用

页码：200 / 202

页数：3

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