Diniz-Sousa Rafaela, Kayano Anderson M, Caldeira Cleópatra A, Simões-Silva Rodrigo, Monteiro Marta C, Moreira-Dill Leandro S, Grabner Fernando P, Calderon Leonardo A, Zuliani Juliana P, Stábeli Rodrigo G, Soares Andreimar M
Center for the Study of Biomolecules Applicable to Health (CEBio), Oswaldo Cruz Foundation - Rondônia (Fiocruz - Rondônia), Porto Velho, RO Brazil.
2Department of Medicine, Federal University of Rondônia (UNIR), Porto Velho, RO Brazil.
J Venom Anim Toxins Incl Trop Dis. 2018 Feb 15;24:5. doi: 10.1186/s40409-018-0143-1. eCollection 2018.
Wasp venoms constitute a molecular reservoir of new pharmacological substances such as peptides and proteins, biological property holders, many of which are yet to be identified. Exploring these sources may lead to the discovery of molecules hitherto unknown. This study describes, for the first time in hymenopteran venoms, the identification of an enzymatically inactive phospholipase A (PLA) from the venom of the social wasp .
venom was fractioned by molecular exclusion and reverse phase chromatography. For the biochemical characterization of the protein, 1D and 2D SDS-PAGE were performed, along with phospholipase activity assays on synthetic substrates, MALDI-TOF mass spectrometry and sequencing by Edman degradation.
The protein, called PocTX, was isolated using two chromatographic steps. Based on the phospholipase activity assay, electrophoresis and mass spectrometry, the protein presented a high degree of purity, with a mass of 13,896.47 Da and a basic pI. After sequencing by the Edman degradation method, it was found that the protein showed a high identity with snake venom PLA homologues.
This is the first report of an enzymatically inactive PLA isolated from wasp venom, similar to snake PLA homologues.
黄蜂毒液构成了新的药理活性物质的分子库，如肽和蛋白质，其中许多生物活性物质尚未被鉴定。探索这些来源可能会发现迄今未知的分子。本研究首次描述了在膜翅目毒液中，从社会性黄蜂毒液中鉴定出一种无酶活性的磷脂酶A（PLA）。
毒液通过分子排阻色谱和反相色谱进行分离。为了对该蛋白质进行生化特性分析，进行了一维和二维SDS-PAGE，同时对合成底物进行磷脂酶活性测定、基质辅助激光解吸电离飞行时间质谱分析以及埃德曼降解测序。
该蛋白质名为PocTX，通过两步色谱法分离得到。基于磷脂酶活性测定、电泳和质谱分析，该蛋白质呈现出高度的纯度，分子量为13,896.47 Da，具有碱性pI值。通过埃德曼降解法测序后发现，该蛋白质与蛇毒PLA同源物具有高度同源性。
这是首次报道从黄蜂毒液中分离出的无酶活性PLA，与蛇PLA同源物相似。
