title: Secretory phospholipase A2 activity in blood serum: the challenge to sense.
abstract: 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.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117997, Russia.
authors.forename: A S
authors.initials: AS
authors.lastname: Alekseeva
authors.affiliation: Russian Cardiology Research and Production Complex, Federal Agency for Health Care, ul. 3ya Cherepkovskaya 15a, Moscow 121552, Russia.
authors.forename: A A
authors.initials: AA
authors.lastname: Korotaeva
authors.affiliation: Russian Cardiology Research and Production Complex, Federal Agency for Health Care, ul. 3ya Cherepkovskaya 15a, Moscow 121552, Russia.
authors.forename: E V
authors.initials: EV
authors.lastname: Samoilova
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117997, Russia.
authors.forename: P E
authors.initials: PE
authors.lastname: Volynsky
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117997, Russia.
authors.forename: E L
authors.initials: EL
authors.lastname: Vodovozova
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117997, Russia; Pirogov Russian National Research Medical University, 1 Ostrovityanov Street, Moscow 117997, Russia. Electronic address: ivan@lipids.ibch.ru.
authors.forename: I A
authors.initials: IA
authors.lastname: Boldyrev
mesh_terms: D015415:Biomarkers / Q000097:blood; D001774:Blood Chemical Analysis / Q000379:methods* / Q000706:statistics & numerical data; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D006868:Hydrolysis; D018836:Inflammation Mediators / Q000097:blood; D007700:Kinetics; D008823:Micelles; D008954:Models, Biological; D054497:Phospholipases A2, Secretory / Q000097:blood*; D010743:Phospholipids / Q000737:chemistry; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
chemical_list: D015415:Biomarkers; D005456:Fluorescent Dyes; D018836:Inflammation Mediators; D008823:Micelles; D010743:Phospholipids; D054497:Phospholipases A2, Secretory
keywords: Fluorescence sensing; Kinetics modeling; Probe design; Secretory phospholipase A2