NDLI: Eukaryotic expression and purification of anti-epilepsy peptide of Buthus martensii Karsch and its protein interactions

Content Provider	Springer Nature Link
Author	Wang, Zongren Wang, Wen Shao, Zhongjun Gao, Bifeng Li, Junchang Ma, Jing Li, Jinghua Che, Honglei Zhang, Wei
Copyright Year	2009
Abstract	The Asian scorpion Buthus martensil Karsch is important in the Chinese traditional medicine where it is used for the treatment of some nervous system diseases. The anti-epilepsy peptide (AEP) is a 61-amino-acid polypeptide extracted from the venom of B. martensil Karsch. Research has confirmed that it has anti-epileptic effects on the rat model of epilepsy. In this experiment, a cDNA library of AEP from the venom of B. martensil Karsch was constructed using RT-PCR; the primer was designed and used for the amplification. An expression vector of AEP was constructed using Pichia pastoris. Vector expression was induced, and protein purification was then performed. Bolting of the interaction molecule of AEP was by His pull down. Experimental results indicate high AEP expression, and the obtained protein was purified and compared with the control group. Four conspicuous protein bands were observed, and mass chromatographic analysis indicated that the four proteins were synaptosomal-associated protein of 25 kDa (SNAP-25), glial fibrillary acidic protein (GFAP), Glutamic acid decarboxylase (GAD) and N-methyl-d-aspartate (NMDA). Further, the four protein bands were verified by mammalian two-hybrid experiments and co-immunoprecipitation. AEP was found to interact with SNAP2 and NMDA. This provides experimental evidence for the mechanism of AEP’s anti-epileptic action and for the manufacture of a novel type anti-epileptic drug.
Starting Page	97
Ending Page	104
Page Count	8
File Format	PDF
ISSN	03008177
Journal	Molecular and Cellular Biochemistry
Volume Number	330
Issue Number	1-2
e-ISSN	15734919
Language	English
Publisher	Springer US
Publisher Date	2009-04-16
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Anti-epilepsy peptide Tandem repeat His pull down Mass chromatographic analysis Cardiology Oncology Medical Biochemistry Biochemistry
Content Type	Text
Resource Type	Article
Subject	Cell Biology Medicine Clinical Biochemistry Molecular Biology

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