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Analytical workflow for rapid screening and purification of bioactives from venom proteomes.

Authors :
Otvos RA
Heus F
Vonk FJ
Halff J
Bruyneel B
Paliukhovich I
Smit AB
Niessen WM
Kool J
Source :
Toxicon : official journal of the International Society on Toxinology [Toxicon] 2013 Dec 15; Vol. 76, pp. 270-81. Date of Electronic Publication: 2013 Oct 16.
Publication Year :
2013

Abstract

Animal venoms are important sources for finding new pharmaceutical lead molecules. We used an analytical platform for initial rapid screening and identification of bioactive compounds from these venoms followed by fast and straightforward LC-MS only guided purification to obtain bioactives for further chemical and biological studies. The analytical platform consists of a nano-LC separation coupled post-column to high-resolution mass spectrometry and parallel on-line bioaffinity profiling for the acetylcholine binding protein (AChBP) in a chip based fluorescent enhancement based bioassay. AChBP is a stable structural homologue of the extracellular ligand binding domain of the α7-nicotinic acetylcholine receptor (α7-nAChR). This receptor is an extensively studied medicinal target, previously associated with epilepsy, Alzheimer's, schizophrenia and anxiety. The workflow is demonstrated with the venom of the Naja mossambica mossambica. Two medium affinity AChBP ligands were found. After subsequent LC-MS guided purification of the respective venom peptides, the purified peptides were sequenced and confirmed as Cytotoxin 1 and 2. These peptides were not reported before to have affinity for the AChBP. The purified peptides can be used for further biological studies.<br /> (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Details

Language :
English
ISSN :
1879-3150
Volume :
76
Database :
MEDLINE
Journal :
Toxicon : official journal of the International Society on Toxinology
Publication Type :
Academic Journal
Accession number :
24140918
Full Text :
https://doi.org/10.1016/j.toxicon.2013.10.013