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- W2897850056 endingPage "e0205756" @default.
- W2897850056 startingPage "e0205756" @default.
- W2897850056 abstract "As a step toward the development of variant forms of Env with enhanced immunogenic properties, we have expressed the glycoprotein in the yeast surface display system in a form that can be subjected to random mutagenesis followed by screening for forms with enhanced binding to germline antibodies. To optimize the expression and immunogenicity of the yeast-displayed Env protein, we tested different approaches for cell wall anchoring, expression of gp120 and gp140 Env from different viral strains, the effects of introducing mutations designed to stabilize Env, and the effects of procedures for altering N-linked glycosylation of Env. We find that diverse forms of HIV envelope glycoprotein can be efficiently expressed at the yeast cell surface and that gp140 forms of Env are effectively cleaved by Kex2p, the yeast furin protease homolog. Multiple yeast-displayed gp120 and gp140 proteins are capable of binding to antibodies directed against the V3-variable loop, CD4 binding site, and gp41 membrane-proximal regions, including some antibodies whose binding is known to depend on Env conformation and N-linked glycan. Based on antibody recognition and sensitivity to glycosidases, yeast glycosylation patterns partially mimic high mannose-type N-glycosylation in mammalian cells. However, yeast-displayed Env is not recognized by some anti-Env antibodies sensitive to quaternary structure, suggesting either that the displayed protein exists in a monomeric state or that for these antibodies, yeast glycosylation in certain regions hinders recognition or access. Consistent with studies in other systems, reconstructed predicted unmutated precursors to anti-Env antibodies exhibit little affinity for the yeast-displayed envelope protein." @default.
- W2897850056 created "2018-10-26" @default.
- W2897850056 creator A5006732358 @default.
- W2897850056 creator A5008068505 @default.
- W2897850056 creator A5029179787 @default.
- W2897850056 creator A5045874910 @default.
- W2897850056 creator A5050355532 @default.
- W2897850056 creator A5054496921 @default.
- W2897850056 creator A5069394146 @default.
- W2897850056 creator A5073227816 @default.
- W2897850056 creator A5086210347 @default.
- W2897850056 date "2018-10-18" @default.
- W2897850056 modified "2023-09-26" @default.
- W2897850056 title "Display of the HIV envelope protein at the yeast cell surface for immunogen development" @default.
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