Secretory phospholipase A2 activity in blood serum: The challenge to sense

被引：12

作者：

Alekseeva, A. S. ^{[1
]}

Korotaeva, A. A. ^{[2
]}

Samoilova, E. V. ^{[2
]}

Volynsky, P. E. ^{[1
]}

Vodovozova, E. L. ^{[1
]}

Boldyrev, I. A. ^{[1
,3
]}

机构：

[1] Russian Acad Sci, Shemyakin Ovchinnikov Inst Bioorgan Chem, Moscow 117997, Russia

[2] Fed Agcy Hlth Care, Russian Cardiol Res & Prod Complex, Moscow 121552, Russia

[3] Pirogov Russian Natl Res Med Univ, Moscow 117997, Russia

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2014年 / 454卷 / 01期

基金：

俄罗斯基础研究基金会;

关键词：

Secretory phospholipase A2; Fluorescence sensing; Probe design; Kinetics modeling; CORONARY EVENTS; A(2) ENZYMES; DISEASE; IIA; FLUORESCENCE; POTENT; ASSAYS;

D O I：

10.1016/j.bbrc.2014.10.069

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：

Excess levels of secretory phospholipase A2 (sPLA2) is known to contribute to several inflammatory diseases including vascular inflammation correlating with coronary events in coronary artery disease. Thus a method to monitor sPLA2 activity in blood serum is urgently needed. Such method is still a challenge since existing fluorescent probes do not allow to monitor sPLA2 activity directly in blood serum. Here we analyze and overcome barriers in sPLA2 sensing methodology and report a fluorescent probe and a kinetic model of its hydrolysis by sPLA2. New probe is designed with a fluorophore and a quencher not interfering binding to the enzyme. At the same time phospholipid matrix bearing the probe promotes efficient initial quenching of the fluorophore. Kinetic model of probe hydrolysis takes into account signal change due to the side processes. The probe and the kinetic model applied together prove the concept that the activity of sPLA can be measured directly in blood serum. (C) 2014 Elsevier Inc. All rights reserved.

引用

页码：178 / 182

页数：5

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