LSTM Home > LSTM Research > LSTM Online Archive

Liquid chromatographic nanofractionation with parallel mass spectrometric detection for the screening of plasmin inhibitors and (metallo)proteinases in snake venoms.

Downloads

Downloads per month over past year

Zietek, Barbara M, Mayar, Morwarid, Slagboom, Julien, Bruyneel, Ben, Vonk, Freek J, Somsen, Govert W, Casewell, Nicholas ORCID: https://orcid.org/0000-0002-8035-4719 and Kool, Jeroen (2018) 'Liquid chromatographic nanofractionation with parallel mass spectrometric detection for the screening of plasmin inhibitors and (metallo)proteinases in snake venoms.'. Analytical and Bioanalytical Chemistry, Vol 410, Issue 23, pp. 5751-5763.

[img]
Preview
Text
analytical & bioanalytical chemistry_Liquid chromatographic nanofractionation.pdf - Published Version
Available under License Creative Commons Attribution.

Download (1MB) | Preview

Abstract

To better understand envenoming and to facilitate the development of new therapies for snakebite victims, rapid, sensitive, and robust methods for assessing the toxicity of individual venom proteins are required. Metalloproteinases comprise a major protein family responsible for many aspects of venom-induced haemotoxicity including coagulopathy, one of the most devastating effects of snake envenomation, and is characterized by fibrinogen depletion. Snake venoms are also known to contain anti-fibrinolytic agents with therapeutic potential, which makes them a good source of new plasmin inhibitors. The protease plasmin degrades fibrin clots, and changes in its activity can lead to life-threatening levels of fibrinolysis. Here, we present a methodology for the screening of plasmin inhibitors in snake venoms and the simultaneous assessment of general venom protease activity. Venom is first chromatographically separated followed by column effluent collection onto a 384-well plate using nanofractionation. Via a post-column split, mass spectrometry (MS) analysis of the effluent is performed in parallel. The nanofractionated venoms are exposed to a plasmin bioassay, and the resulting bioassay activity chromatograms are correlated to the MS data. To study observed proteolytic activity of venoms in more detail, venom fractions were exposed to variants of the plasmin bioassay in which the assay mixture was enriched with zinc or calcium ions, or the chelating agents EDTA or 1,10-phenanthroline were added. The plasmin activity screening system was applied to snake venoms and successfully detected compounds exhibiting antiplasmin (anti-fibrinolytic) activities in the venom of Daboia russelii, and metal-dependent proteases in the venom of Crotalus basiliscus. Graphical abstract ᅟ.

Item Type: Article
Subjects: QV Pharmacology > Hematologic Agents > QV 195 Hemostatics. Coagulants
QV Pharmacology > Toxicology > General Toxicology > QV 602 Detection of poisons. Tests. Laboratory manuals. Technique
QY Clinical Pathology > QY 25 Laboratory techniques and procedure
WD Disorders of Systemic, Metabolic or Environmental Origin, etc > Animal Poisons > WD 410 Reptiles
Faculty: Department: Biological Sciences > Department of Tropical Disease Biology
Digital Object Identifer (DOI): https://doi.org/10.1007/s00216-018-1253-x
Depositing User: Stacy Murtagh
Date Deposited: 14 Aug 2018 10:28
Last Modified: 16 Aug 2018 12:42
URI: http://archive.lstmed.ac.uk/id/eprint/9061

Statistics

View details

Actions (login required)

Edit Item Edit Item