Characterization of a Trypanosoma brucei Alkb homolog capable of repairing alkylated DNA

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• 作者：Jana M. Simmons^a ; ¹ ; Donna J. Koslowsky^b ; Robert P. Hausinger^a ; ^b ; ^{hausinge@msu.edu}
• 关键词：伪KG ; 伪-ketoglutarate ; MMS ; methyl methanesulfonate ; ODPA ; ortho-phenylenediamine ; TbABH ; AlkB homolog from Trypanosoma brucei
• 刊名：Experimental Parasitology
• 出版年：2012
• 期刊代码：99_00144894
• 类别：imm
• 出版时间：May, 2012
• 卷：131
• 期：1
• 页码：92-100
• 文件大小：1220 K

摘要

Trypanosoma brucei encodes a protein (denoted TbABH) that is homologous to AlkB of Escherichia coli and AlkB homolog (ABH) proteins in other organisms, raising the possibility that trypanosomes catalyze oxidative repair of alkylation-damaged DNA. TbABH was cloned and expressed in E. coli, and the recombinant protein was purified and characterized. Incubation of anaerobic TbABH with Fe^II and 伪-ketoglutarate (伪KG) produces a characteristic metal-to-ligand charge-transfer chromophore, confirming its membership in the Fe^II/伪KG dioxygenase superfamily. The protein binds to DNA, with a clear preference for alkylated oligonucleotides according to results derived by electrophoretic mobility shift assays. Finally, the protozoan gene was shown to partially complement E. coli alkB cells when stressed with methylmethanesulfonate; thus confirming assignment of TbABH as a functional AlkB protein in T. brucei.