Methods development for Analysis of Partially Deglycosylated Proteins and Application to an HIV Envelope Protein Vaccine Candidate
| Autor: | Ben-Jiang Ma, Eden P. Go, Geetha S. Hewawasam, Hua-Xin Liao, Heather Desaire, Barton F. Haynes |
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| Rok vydání: | 2011 |
| Předmět: |
PNGase F
chemistry.chemical_classification Glycosylation biology Human immunodeficiency virus (HIV) HIV envelope protein macromolecular substances Condensed Matter Physics medicine.disease_cause Endoglycosidase Article carbohydrates (lipids) chemistry.chemical_compound Biochemistry chemistry Site occupancy biology.protein medicine lipids (amino acids peptides and proteins) Physical and Theoretical Chemistry HIV vaccine Glycoprotein Instrumentation Spectroscopy |
| Zdroj: | International journal of mass spectrometry. 305(2-3) |
| ISSN: | 1387-3806 |
| Popis: | The work presented herein describes the first comprehensive analysis of a partially deglycosylated HIV vaccine candidate envelope protein (Env). The Env, JRFL gp140 ΔCF, with 27 potential glycosylation sites, was partially deglycosylated with PNGase F as part of a strategy to generate a more immunogenic HIV vaccine, and the resulting protein's glycosylation was characterized in a unique workflow using two different glycosidases, Endo H and Endo F3. This unique analysis protocol provided for coverage on 26 of the 27 glycosylation sites, and the data showed that the biochemical treatment with PNGase F resulted in a highly heterogeneous glycoprotein product that had been partially deglycosylated at most of the glycosylation sites. The protocols described in this work could be useful for characterizing the glycosylation site occupancy of other native or biochemically deglycosylated proteins. |
| Databáze: | OpenAIRE |
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