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Analysis and characterization of antimicrobial peptides by capillary electromigration methods

SYSNO ASEP	0538687
Document Type	A - Abstract
R&D Document Type	The record was not marked in the RIV
R&D Document Type	Není vybrán druh dokumentu
Title	Analysis and characterization of antimicrobial peptides by capillary electromigration methods
Author(s)	Kašička, Václav (UOCHB-X)_{RID, ORCID} Tůmová, Tereza (UOCHB-X)_RID Monincová, Lenka (UOCHB-X) Čeřovský, Václav (UOCHB-X)_{RID, ORCID}
Source Title	Czech Chemical Society Symposium Series - ISSN 2336-7202 Roč. 18, č. 3 (2020), s. 68
Number of pages	1 s.
Publication form	Print - P
Action	Sjezd českých a slovenských chemických společností /72./
Event date	06.09.2020 - 09.09.2020
VEvent location	Praha
Country	CZ - Czech Republic
Event type	EUR
Language	eng - English
Country	CZ - Czech Republic
Keywords	antimicrobial peptides ; capillary electrophoresis ; capillary isotachophoresis
Subject RIV	CB - Analytical Chemistry, Separation
OECD category	Analytical chemistry
R&D Projects	GA18-02597S GA ČR - Czech Science Foundation (CSF)
Institutional support	UOCHB-X - RVO:61388963
Annotation	High-performance capillary electromigration methods, zone electrophoresis (CZE) and isotachophoresis (CITP), have been employed for qualitative and quantitative analysis and physicochemical characterization of polycationic antimicrobial peptides (AMPs) isolated from the venom of two species of Hymenoptera insects. These AMPs are composed of 10–20 amino acids (AAs) with 3 to 7 basic AAs (His, Lys, Arg). Purity degree of synthetic AMPs was quantified by CZE in acidic background electrolytes (BGEs) using capillary coatings with cationic surfactants or polymers suppressing peptide sorption to the capillary wall. Acid-base and electromigration properties of the AMPs were investigated by combination of CZE and CITP. First, the mixed acidity constants, pKa,mix, and the actual ionic mobilities of the AMPs were estimated by nonlinear regression analysis of the pH dependence of their effective mobilities measured in series of BGEs within wide pH range (1.80–12.10), at constant ionic strength (25 mM) and temperature (25 °C). Then, the pKa,mix values were recalculated to the thermodynamic pKas using the DebyeHückel equation for activity coefficients. Effective charge of the AMPs was estimated by CITP. This approach is based on the linear dependence of the CITP zone length of the analyte on its effective charge and injected substance amount. Cationic CITP systems with leading electrolyte (LE) composed of 10 mM ammonium leading ion, 40 mM acetate counterion, pH 4.1, and terminating electrolyte containing 40 mM acetic acid, pH 3.2, provided sharp AMPs zones with good linearity and repeatability of their lengths. Ionic mobilities of AMPs were determined by CZE in the BGE of the same composition as the LE. Using this procedure, the effective charge numbers of the AMPs were found to be in the range 1.65–5.04, i.e. much less than the theoretical charge numbers (2.86–6.99) calculated from the pKa values.
Workplace	Institute of Organic Chemistry and Biochemistry
Contact	asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434
Year of Publishing	2021

Number of the records: 1