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利用磷光和光探测磁共振研究1型人类免疫缺陷病毒核衣壳蛋白与核酸的结合

Binding of the nucleocapsid protein of type 1 human immunodeficiency virus to nucleic acids studied using phosphorescence and optically detected magnetic resonance.

作者信息

Wu J Q, Ozarowski A, Maki A H, Urbaneja M A, Henderson L E, Casas-Finet J R

机构信息

Department of Chemistry, University of California, Davis, California 95616, USA.

出版信息

Biochemistry. 1997 Oct 14;36(41):12506-18. doi: 10.1021/bi970676f.

DOI:10.1021/bi970676f
PMID:9376355
Abstract

The binding of p7 nucleocapsid protein of type 1 human immunodeficiency virus (HIV-1) to various oligonucleotides and polynucleotides has been investigated by phosphorescence and optically detected magnetic resonance (ODMR) spectroscopy. The intrinsic spectroscopic probe used in these studies is the photoexcited triplet state of Trp37, which is associated with the C-terminal zinc finger of p7 and is its only tryptophan residue. Complex formation produces a red-shift of the phosphorescence 0, 0-band (DeltaE0,0) of Trp37 as well as a reduction of the zero field splitting (zfs) D parameter. Increases of -DeltaE0,0 (A < C < U < G <I) rank with increasing binding affinity to nucleic acid homooligomers (A approximately C < U < G approximately I). It is proposed that the magnitude of the shift reflects the extent of aromatic stacking interactions. We propose also that -DeltaD increases not only with increased aromatic stacking but also with the extent of charge transfer (CT) character admixed into the triplet state. The quantity DeltaD/DeltaE0,0 correlates with the electron affinity of the bases (G < A < C < U approximately T), suggesting that this quantity reflects the extent of CT character admixed with the triplet state by the aromatic stacking interaction. Also affected by nucleic acid binding of p7 are the kinetic parameters of Trp37. We find a selective increase in the relative populating rate, and of the decay rate constant of the Tx sublevel. In binding of p7 to either d(IT)2 or d(IT)4, two distinct sets of triplet states of Trp37 are resolved, suggesting the existence of specific nucleic acid binding modes of these heterooligomers.

摘要

通过磷光和光探测磁共振(ODMR)光谱研究了1型人类免疫缺陷病毒(HIV-1)的p7核衣壳蛋白与各种寡核苷酸和多核苷酸的结合。这些研究中使用的内在光谱探针是Trp37的光激发三重态,它与p7的C末端锌指相关,并且是其唯一的色氨酸残基。复合物的形成导致Trp37的磷光0,0带(ΔE0,0)发生红移,同时零场分裂(zfs)D参数减小。-ΔE0,0的增加(A < C < U < G <I)与对核酸同聚体的结合亲和力增加(A≈C < U < G≈I)呈正相关。有人提出,这种位移的大小反映了芳香堆积相互作用的程度。我们还提出,-ΔD不仅随着芳香堆积的增加而增加,而且随着混入三重态的电荷转移(CT)特征的程度而增加。ΔD/ΔE0,0的量与碱基的电子亲和力相关(G < A < C < U≈T),这表明该量反映了通过芳香堆积相互作用混入三重态的CT特征的程度。Trp37的动力学参数也受p7核酸结合的影响。我们发现相对填充率和Tx子能级的衰减速率常数有选择性增加。在p7与d(IT)2或d(IT)4的结合中,分辨出Trp37的两组不同的三重态,这表明这些异聚体存在特定的核酸结合模式。

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