A non-cleavable hexahistidine affinity tag at the carboxyl-terminus of the HIV-1 Pr55^Gag polyprotein alters nucleic acid binding properties

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• 作者：Maria C. Bewley^a ; Lisa Reinhart^a ; Matthew S. Stake^b ; ^c ; Shorena Nadaraia-Hoke^b ; ^c ; Leslie J. Parent^b ; ^c ; ; John M. Flanagan^a ; ^{jmf27@psu.edu}
• 关键词：HIV ; Gag ; Pr55Gag ; Nucleic acid binding ; p6 ; Proteolysis ; His-tag
• 刊名：Protein Expression and Purification
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：130
• 期：Complete
• 页码：137-145
• 全文大小：1168 K
• 卷排序：130

文摘

The effect on nucleic acid binding of hexahistidine tags fused the C-terminus of full length HIV Pr55Gag was investigated. A standard expression and purification protocol minimized the differences in trace metal levels and protein contaminants. The hexahistidine tagged Pr55Gagbinds nucleic acids more tightly than untagged Pr55Gag and has altered binding behavior. Removal of the hexahistidine tag resulted in binding curves similar to untagged Pr55Gag. A cleavable His tag should be considered when isolating Pr55Gag.