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Comparative Study
. 1994 Nov 7;354(2):200-2.
doi: 10.1016/0014-5793(94)01110-9.

Affinity Modification of Human Immunodeficiency Virus Reverse Transcriptase and DNA Template by Photoreactive dCTP Analogs

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Comparative Study

Affinity Modification of Human Immunodeficiency Virus Reverse Transcriptase and DNA Template by Photoreactive dCTP Analogs

S V Doronin et al. FEBS Lett. .
Free article

Abstract

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' 32P end-labeled 3'-(d)TTACATATCGGGGATGGT-5' primer were added. The 5' 32P 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.

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