Recombinant human O 6‐alkylguanine‐DNA alkyltransferase induces conformational change in bound DNA

FEBS Letters

Bender, Kirsten²  Hassiepen, Ulrich¹  Wollmer, Axel¹  Federwisch, Matthias²  Rajewsky, Manfred F² 

[1] Institute of Biochemistry, Rheinisch-Westfälische Technische Hochschule, Pauwelsstrasse 30, D-52057 Aachen, Germany;Institute of Cell Biology (Cancer Research) [IFZ], University of Essen Medical School, Hufeland-Strasse 55, D-45122 Essen, Germany

关键词: O 6-Alkylguanine-DNA alkyltransferase;    Protein-DNA interaction;    Circular dichroism;    Time-resolved fluorescence;    AGT;    O 6-alkylguanine-DNA alkyltransferase;    DTT;    1;    4-dithiothreitol;    FAD;    fluorescence anisotropy decay;    FWHM;    full width at half maximum;    NTA;    nitrilo-triacetic acid;   

DOI  :  10.1016/S0014-5793(97)00370-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Circular dichroism, and steady-state and time-resolved fluorescence spectroscopy were used to compare the native recombinant human DNA repair protein O ⁶-alkylguanine-DNA alkyltransferase (AGT) with AGT bound to ds-DNA. Contrary to fluorescence, analysis of the far-UV CD spectra indicated a conformational change of AGT upon binding to DNA: its α-helical content is increased by ∼12%. Analysis of near-UV CD spectra revealed that DNA was also affected, probably being separated into single strands locally.

【 授权许可】

Unknown