Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6492225 | Journal of Biotechnology | 2011 | 8 Pages |
Abstract
We demonstrated a high level expression and purification of recombinant human immunodeficiency virus type 1 gp41 ectodomain (gp41e-FP) using glass bead approach with a final yield of 12 ± 2 mg/L bacterial culture. The proper folding of gp41e-FP encompassing the fusion peptide (FP) was ascertained by circular dichroism (CD) measurement and recognition by NC-1 antibody. The latter assay revealed stabilization of the gp41 coiled coil structure in the presence of liposome dispersion. The differential affinity of gp41e-FP and gp41e (devoid of FP) by NC-1 suggested an aggregated state for gp41e-FP and/or possible proximity of the fusion peptide domain to the coiled coil structure of gp41 ectodomain. Perfluorooctanoate (PFO)-PAGE electrophoresis experiment revealed the trimeric propensity of the recombinant gp41e-FP. In comparison to gp41e, the lipid mixing activity of gp41e-FP was two-fold higher suggesting a role of FP in promoting membrane fusion. The present approach to efficiently and quantitatively preparing the functional full-length recombinant gp41 ectodomain protein can be employed for structural and biomedical investigations and the extraction of other inclusion body-embedded recombinant proteins.
Keywords
mAbRho-PEDMPGNBD-PEPFOdMPCDPPCGSTMBP1,2-Dimyristoyl-sn-glycero-3-phosphoglycerol1,2-dimyristoyl-sn-glycero-3-phosphocholineMonoclonal antibodyShbSix-helix bundleSurface plasmon resonanceSPRProtein purificationSelf-assemblycircular dichroismPerfluorooctanoateLipid mixingHIV-1human immunodeficiency virusmaltose binding proteinFusion peptideglutathione S-transferase
Related Topics
Physical Sciences and Engineering
Chemical Engineering
Bioengineering
Authors
Chun-Hung Lin, Chi-Hui Lin, Chung-Chieh Chang, Ting-Shyang Wei, Shu-Fang Cheng, Steve S.-L. Chen, Ding-Kwo Chang,