Heterologous expression and functional characterization of phytaspase, a caspase-like plant protease

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• 作者：Sharmila Narayanan^a ; Pallab Sanpui^b ; Lingaraj Sahoo^a ; Siddhartha Sankar Ghosh^a ; ^b ; ^{sghosh@iitg.ernet.in}
• 关键词：Programmed cell death ; Caspase ; Nicotiana tabacum ; Phytaspase ; Serine proteases ; Molecular docking
• 刊名：International Journal of Biological Macromolecules
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：95
• 期：Complete
• 页码：288-293
• 全文大小：1569 K
• 卷排序：95

文摘

First report of successful cloning and expression of tobacco phytaspase gene in Escherichia coli BL21. Structural analysis revealed recombinant phytaspase as α + β class of protein and similarity with plant serine-proteases. Recombinant phytaspase demonstrated caspase-like functional activity. KM and kcat/KM of phytaspase for hydrolyzing Ac-VETD-AMC were 1.587 μM and 4.67 × 103 M−1 min−1, respectively. Transient expression of phytaspase in A549 cancer cells resulted in reduced IC50 value of doxorubicin.