Comparison of the Specificity of Interaction of Cellular and Viral Zinc-Binding Domains with 2-Mercaptobenzamide Thioesters
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- 作者：Lisa M. Miller Jenkins ; Stewart R. Durell ; Andrew T. Maynard ; Stephen J. Stahl ; John K. Inman ; Ettore Appella ; Pascale Legault ; James G. Omichinski
- 刊名：Journal of the American Chemical Society
- 出版年：2006
- 出版时间：September 13, 2006
- 年：2006
- 卷：128
- 期：36
- 页码：11964 - 11976
- 全文大小：382K
- 年卷期：v.128,no.36(September 13, 2006)
- ISSN：1520-5126
文摘
The interactions of two 2-mercaptobenzamide thioester compounds with six diverse zinc-bindingdomains (ZBDs) have been analyzed by UV/visible spectroscopy, NMR spectroscopy, and nucleic acidbinding assays. These thioester compounds serve as useful tools for probing the intrinsic chemical stabilityof ZBDs that exist within a variety of cellular and viral proteins. In our studies, the classical (Cys2His2) zincfinger ZBDs, the interleaved RING like ZBDs of protein kinase C 
(Cys2HisCys and HisCys3), and thecarboxyl-terminal (Cys2HisCys) ZBD of Mouse Mammary Tumor Virus nucleocapsid protein (MMTV NCp10)were resistant to reaction with the thioester compounds. In contrast, the thioester compounds were able toefficiently eject zinc from the amino-terminal (Cys2HisCys) ZBD of MMTV NCp10, a Cys2HisCys ZBD fromFriend of GATA-1 (FOG-1), and from both Cys4 ZBDs of GATA-1. In all cases, zinc ejection led to a lossof protein structure. Interestingly, GATA-1 was resistant to reaction with the thioester compounds whenbound to its target DNA sequence. The electronic and steric screening was calculated for select ZBDs tofurther explore their reactivity. Based on these results, it appears that both first and second zinc-coordinationshell interactions within ZBDs, as well as nucleic acid binding, play important roles in determining thechemical stability and reactivity of ZBDs. These studies not only provide information regarding the relativereactivity of cysteine residues within structural ZBDs but also are crucial for the design of future therapeuticagents that selectively target ZBDs, such as those that occur in the HIV-1 nucleocapsid protein.
(Cys2HisCys and HisCys3), and thecarboxyl-terminal (Cys2HisCys) ZBD of Mouse Mammary Tumor Virus nucleocapsid protein (MMTV NCp10)were resistant to reaction with the thioester compounds. In contrast, the thioester compounds were able toefficiently eject zinc from the amino-terminal (Cys2HisCys) ZBD of MMTV NCp10, a Cys2HisCys ZBD fromFriend of GATA-1 (FOG-1), and from both Cys4 ZBDs of GATA-1. In all cases, zinc ejection led to a lossof protein structure. Interestingly, GATA-1 was resistant to reaction with the thioester compounds whenbound to its target DNA sequence. The electronic and steric screening was calculated for select ZBDs tofurther explore their reactivity. Based on these results, it appears that both first and second zinc-coordinationshell interactions within ZBDs, as well as nucleic acid binding, play important roles in determining thechemical stability and reactivity of ZBDs. These studies not only provide information regarding the relativereactivity of cysteine residues within structural ZBDs but also are crucial for the design of future therapeuticagents that selectively target ZBDs, such as those that occur in the HIV-1 nucleocapsid protein.