Investigating the interfacial binding of bacterial phosphatidylinositol-specific phospholipase C

被引:18
作者
Wehbi, H
Feng, JW
Kolbeck, J
Ananthanarayanan, B
Cho, W
Roberts, MF [1 ]
机构
[1] Boston Coll, Merkert Chem Ctr, Chestnut Hill, MA 02467 USA
[2] Univ Illinois, Dept Chem, Chicago, IL 60607 USA
关键词
D O I
10.1021/bi034195+
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The interactions of PI-PLC with nonsubstrate zwitterionic [phosphatidylcholine (PC)] and anionic [phosphatidylmethanol (PMe), phosphatidylserine, phosphatidylglycerol, and phosphatidic acid] interfaces that affect the catalytic activity of PI-PLC have been examined. PI-PLC binding is strongly coupled to vesicle curvature and is tighter at acidic pH for all of the phospholipids examined. PI-PLC binds to small unilamellar vesicles (SUVs) of anionic lipids with much higher affinity (K-d is 0.01-0.07 muM for a site consisting of n = 100 +/- 25 lipids when analyzed with a Langmuir adsorption isotherm) than to zwitterionic PC SUVs (K-d is 5-20 muM and n = 8 +/- 3). The binding to PC surfaces is dominated by hydrophobic interactions, while binding to anionic surfaces is dominated by electrostatic interactions. The contributions of specific cationic side chains and hydrophobic groups at the rim of the alphabeta-barrel to zwitterionic and anionic vesicle binding have been assessed with mutagenesis. The results are used to explain how PC activates the enzyme for both phosphotransferase and cyclic phosphodiesterase activities.
引用
收藏
页码:9374 / 9382
页数:9
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