Purification, characterization and gene cloning of a novel phospholipase A2 from the venom of Agkistrodon blomhoffii ussurensis |
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Authors: | Bao Yongming Bu Pengcheng Jin Liji Hongxia Wang Yang Qing An Lijia |
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Affiliation: | Department of Bioscience and Biotechnology, Dalian University of Technology, 2 Linggong Road, Dalian 116024, PR China. yongmingbao@sohu.com |
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Abstract: | A new phospholipase A2 with Gln at the site 49, abbreviated as Gln49-PLA2, has been purified from the venom of Agkistrodon blomhoffii ussurensis by using ion-exchange chromatography, gel filtration chromatography and reversed-phase HPLC, and behaves as a single-band on SDS-PAGE. Its molecular weight is 13881.85+/-0.33 Da given by mass spectrometry and pI is about 8.56 given by isoelectric focusing. Gln49-PLA2 does not show phospholipase A2 and hemorrhagic activity, whereas shows weak toxic and apparent anticoagulant activity. Based on the N-terminal sequencing and peptide mass fingerprint analysis, Gln49-PLA2 cDNA has been cloned by means of RT-PCR. Gln49-PLA2 consists of 122 amino acid residues and has the structural features of class II of snake venom phospholipase A2. |
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