Site-Selective Protein Immobilization by Covalent Modification of GST Fusion Proteins

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• 作者：Yiqing Zhou ; Tianlin Guo ; Guanghui Tang ; Hui Wu ; Nai-Kei Wong ; Zhengying Pan
• 刊名：Bioconjugate Chemistry
• 出版年：2014
• 出版时间：November 19, 2014
• 年：2014
• 卷：25
• 期：11
• 页码：1911-1915
• 全文大小：272K
• ISSN：1520-4812

文摘

The immobilization of functional proteins onto solid supports using affinity tags is an attractive approach in recent development of protein microarray technologies. Among the commonly used fusion protein tags, glutathione S-transferase (GST) proteins have been indispensable tools for protein鈥損rotein interaction studies and have extensive applications in recombinant protein purification and reversible protein immobilization. Here, by utilizing pyrimidine-based small-molecule probes with a sulfonyl fluoride reactive group, we report a novel and general approach for site-selective immobilization of Schistosoma japonicum GST (sjGST) fusion proteins through irreversible and specific covalent modification of the tyrosine-111 residue of the sjGST tag. As demonstrated by sjGST-tagged eGFP and sjGST-tagged kinase activity assays, this immobilization approach offers the advantages of high immobilization efficiency and excellent retention of protein structure and activity.