Listeria monocytogenes phosphatidylinositol-specific phospholipase C: activation and allostery

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• 作者：Ryan ; Margret ; Zaikova ; Tatiana O. ; Keana ; John F.W. ; Goldfine ; Howard ; Griffith ; O. Hayes
• 关键词：Enzyme kinetics ; Interfacial activation ; Phosphatidylinositol-specific phospholipase C (PI-PLC) ; Co-operativity
• 刊名：Biophysical Chemistry
• 出版年：2002
• 出版时间：December 10, 2002
• 年：2002
• 卷：101-102
• 期：Complete
• 页码：347-358
• 全文大小：197 K

文摘

The animal and human pathogen Listeria monocytogenes secretes several virulence factors, including a phosphatidylinositol-specific phospholipase C (PI-PLC). Sufficient quantities of L. monocytogenes PI-PLC for biophysical studies were obtained by overexpression of the enzyme in Escherichia coli. The purified PI-PLC was examined in enzyme kinetics experiments using a new fluorogenic substrate, methyl-FLIP. Methyl-FLIP is a water-soluble monomeric substrate cleaved in a manner similar to the natural aggregate substrate, phosphatidylinositol (PI). Michaelis–Menten kinetics were observed with K_M