Identification and Characterization of Heparan Sulphate Binding Proteins of <i>Entamoeba histolytica</i>

• 作者: Mohan Lal Dubey Rakesh Sehgal Sumeeta Khurana Uma Nahar Saikia Upninder Kaur
• 来源期刊: 生物科学与医学(英文)
• 学科: 医学
• 关键词: Heparan Sulphate ENTAMOEBA HISTOLYTICA IDENTIFICATION CHARACTERIZATION
• 年，卷（期）: 2018,（8）
• 页码范围: 60-71
• 页数: 12页
• 分类号: R73

摘要

A large number of microbial pathogens bind to heparan sulphate on eukaryotic cell surfaces. Heparan Sulphate Binding Proteins (HSBPs) from Entamoeba histolytica culture lysates were obtained by sequential ammonium sulphate precipitation and Protein purify. SDS-PAGE and immunoblotting experiments indicated the presence of two major extracellular proteins in E. histolytica (51.2 kDa and 61.0 kDa). Characterization of HSBPs by 2D Gel electrophoresis of 40% (NH4)2SO4 precipitated lysate of E. histolytica revealed that the isoelectric point of 51.2 kDa HSBP was at pH3.0. The protein of 61.0 kDa HSBP showed three spots in 40% (NH4)2SO4 lysate which had isoelectric point between pH 4.0 - 7.0. While in 80% (NH4)2SO4 precipitated lysate, 51.2 kDa HSBP showed only one spot which had isoelectric point at pH 3. However, 61.0 kDa HSBP revealed 2 spots which had isoelectric point between pH 4 and 5. The result showed that this parasite has proteins which interact with heparan sulphate whose molecular formula is C14H23NO21S-23. These proteins may have a role in binding of parasite to heparan sulphate on host cells. Further characterization by MALDI-TOF analysis of HSBPs from E. histolytica demonstrated HSBPs to be novel protein in this parasite which has been isolated, purified and characterized first time by our group in the present study.