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Evidence for stacking interactions between 5‐mercurated polyuridylic acid and HIV‐1 p7 nucleocapsid protein obtained by phosphorescence and optically detected magnetic resonance (ODMR)
[摘要]

The photoexcited triplet state of Trp-37 in the C-terminal zinc finger of the HIV-1 p7 nucleocapsid protein was used as a probe of p7 interactions with the heavy atom-derivatized RNA homopolymer, poly-5-mercuriuridylic acid (5-HgU). Binding of p7 to 5-HgU (Hg blocked with 2-mercaptoethanol) produces an external heavy atom effect (HAE) on Trp-37 characterized by fluorescence quenching, reduction of the phosphorescence lifetime by three orders of magnitude, and the appearance of the D + E phosphorescence-detected ODMR signal, absent in unperturbed Trp, but induced by a HAE. The details of the HAE are consistent with out-of-plane van der Waals contact of Hg with the indole chromophore of Trp-37. Steric requirements suggest further that the Trp-RNA contact occurs via an aromatic stacking interaction.

[发布日期]  [发布机构] 
[效力级别]  [学科分类] 生物化学/生物物理
[关键词] External heavy atom effect;Tryptophan phosphorescence;ODMR spectroscopy;Zinc finger-RNA interaction;5-HgU;5-mercurated polyuridylic acid;HIV-1;human immunodeficiency virus type-1;HAE;heavy atom effect;ME;2-mercaptoethanol;MIDP;microwave-induced delayed phosphorescence;NC;nucleocapsid;ODMR;optically detected magnetic resonance of the photoexcited triplet state;SLR;spin-lattice relaxation;SSB;E. coli single-stranded DNA-binding protein;ZFS;zero-field splittings [时效性] 
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