Glycoproteomic study on human immunodeficiency virus infection related glycoproteins / 中华传染病杂志

ABSTRACT

Objective To investigate human plasma glycoproteomie changes related to human immunodeficiency virus (HIV) infection,and to identify glycoproteins with potential anti-HIV activity or anti-HIV drug targets. Methods Plasma proteins with lower abundance were enriched through affinity purification to remove albumin and IgG in clinical samples (HIV-positive patient, n= 10, and healthy controls, n= 20). Proteins were separated by two-dimensional electrophoresis (2-DE) and stained by Pro-Q emerald glycoprotein stain kits. The 2-DE image was analyzed by ImageMaster software to find differential glycoproteins. Furthermore, the depleted HIV-positive and healthy control plasma proteins were digested by PNGase F. Glycoproteins were deglycoliszed, separated by 2-DE and analyzed by ImageMaster software. Differential glycoproteins were identified by liquid chromatography combined with high capacity ion trap mass spectrometry (HCT). Results The pretreatment of HIV-positive plasma prior to 2-DE could efficiently remove the high aboundant albumin and IgG in plasma and improve the detection of proteins with low-abundance. High revolution 2-DE gel images of glycoproteins from HIV positive and healthy control plasma samples were obtained. Glycoproteins were successfully deglycolized through PNGase F treatment. Thirteen differential glycoproteins were identified by liquid chromatography combined with mass spectrometry. These proteins included alphalantitrypsin precursor and serine/threonine-protein kinase N1. Conclusions Potential HIV infection related proteins,such as alphal-antitrypsin precursor are successfully identified. Our study may offer some help to understand the molecular mechanism of HIV infection and select new drug targets for preventing HIV infection.