Lag-burst kinetics in phospholipase A2 hydrolysis of DPPC bilayers visualized by atomic force microscopy

被引:93
作者
Nielsen, LK
Risbo, J
Callisen, TH
Bjornholm, T
机构
[1] Univ Copenhagen, Dept Chem, Labs Mat Sci, CISMI, DK-2100 Copenhagen O, Denmark
[2] Tech Univ Denmark, Dept Chem, DK-2800 Lyngby, Denmark
来源
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | 1999年 / 1420卷 / 1-2期
关键词
lag burst; atomic force microscopy; phospholipase A(2); supported bilayer; enzyme kinetics; nano chemistry;
D O I
10.1016/S0005-2736(99)00103-0
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 lag-burst phenomenon in the phospholipase A(2) mediated hydrolysis of phospholipid bilayers is for the first time demonstrated in an atomic force microscopy (AFM) study. Simultaneous AFM measurements of the degree of bilayer degradation and the physical-chemical state of the membrane reveals growing nanoscale indentations in the membrane during the lag phase. It is argued that these indentations are domains of hydrolysis products (lysoPC/PC) which eventually trigger the burst. The rate of the rapid hydrolysis following the burst is found to be proportional to the length of the edge between membrane adsorbed and desorbed to the mica base. The observed maximal rate of membrane degradation is approx. 0.2 mmol lipid/min/mol lipase in solution. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:266 / 271
页数:6
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