Molecular Characterization of Three Novel Phospholipase A2 Proteins from the Venom of Atheris chlorechis, Atheris nitschei and Atheris squamigera


Autoria(s): Wang, He; Chen, Xiaole; Zhou, Mei; Wang, Lei; Chen, Tianbao; Shaw, Chris
Data(s)

01/06/2016

Resumo

Secretory phospholipase A2 (sPLA2) is known as a major component of snake venoms and displays higher-order catalytic hydrolysis functions as well as a wide range of pathological effects. Atheris is not a notoriously dangerous genus of snakes although there are some reports of fatal cases after envenomation due to the effects of coagulation disturbances and hemorrhaging. Molecular characterization of Atheris venom enzymes is incomplete and there are only a few reports in the literature. Here, we report, for the first time, the cloning and characterization of three novel cDNAs encoding phospholipase A2 precursors (one each) from the venoms of the Western bush viper (Atheris chlorechis), the Great Lakes bush viper (Atheris nitschei) and the Variable bush viper (Atheris squamigera), using a “shotgun cloning” strategy. Open-reading frames of respective cloned cDNAs contained putative 16 residue signal peptides and mature proteins composed of 121 to 123 amino acid residues. Alignment of mature protein sequences revealed high degrees of structural conservation and identity with Group II venom PLA2 proteins from other taxa within the Viperidae. Reverse-phase High Performance Liquid Chromatography (HPLC) profiles of these three snake venoms were obtained separately and chromatographic fractions were assessed for phospholipase activity using an egg yolk suspension assay. The molecular masses of mature proteins were all identified as approximately 14 kDa. Mass spectrometric analyses of the fractionated oligopeptides arising from tryptic digestion of intact venom proteins, was performed for further structural characterization.

Formato

application/pdf

Identificador

http://pure.qub.ac.uk/portal/en/publications/molecular-characterization-of-three-novel-phospholipase-a2-proteins-from-the-venom-of-atheris-chlorechis-atheris-nitschei-and-atheris-squamigera(a6db0fe1-56b7-4551-ad18-9eea761e882f).html

http://dx.doi.org/10.3390/toxins8060168

http://pure.qub.ac.uk/ws/files/47040575/Molecular_Characterization.pdf

Idioma(s)

eng

Direitos

info:eu-repo/semantics/openAccess

Fonte

Wang , H , Chen , X , Zhou , M , Wang , L , Chen , T & Shaw , C 2016 , ' Molecular Characterization of Three Novel Phospholipase A2 Proteins from the Venom of Atheris chlorechis, Atheris nitschei and Atheris squamigera ' Toxins , vol 8 , no. 6 , 168 . DOI: 10.3390/toxins8060168

Palavras-Chave #snake venom; phospholipase A2; molecular cloning; Atheris; mass spectrometry
Tipo

article