Structure of the S. aureus PI-Specific Phospholipase C Reveals Modulation of Active Site Access by a Titratable 蟺-Cation Latched Loop

详细信息    查看全文

• 作者：Rebecca Goldstein ; Jiongjia Cheng ; Boguslaw Stec ; Mary F. Roberts
• 刊名：Biochemistry
• 出版年：2012
• 出版时间：March 27, 2012
• 年：2012
• 卷：51
• 期：12
• 页码：2579-2587
• 全文大小：532K
• 年卷期：v.51,no.12(March 27, 2012)
• ISSN：1520-4995

文摘

Staphylococcus aureus secretes a phosphatidylinositol-specific phospholipase C (PI-PLC) as a virulence factor that is unusual in exhibiting higher activity at acidic pH values than other enzymes in this class. We have determined the crystal structure of this enzyme at pH 4.6 and pH 7.5. Under slightly basic conditions, the S. aureus PI-PLC structure closely follows the conformation of other bacterial PI-PLCs. However, when crystallized under acidic conditions, a large section of mobile loop at the 伪尾-barrel rim in the vicinity of the active site shows 10 脜 shift. This loop displacement at acidic pH is the result of a titratable intramolecular 蟺-cation interaction between His258 and Phe249. This was verified by a structure of the mutant protein H258Y crystallized at pH 4.6, which does not exhibit the large loop shift. The intramolecular 蟺-cation interaction for S. aureus PI-PLC provides an explanation for the activity of the enzyme at acid pH and also suggests how phosphatidylcholine, as a competitor for Phe249, may kinetically activate this enzyme.