Journal of Antivirals & Antiretrovirals
Open Access
ISSN: 1948-5964
ISSN: 1948-5964
Agnes Hajduczki
Posters: JAA
Displaying proteins and peptides on the surface of viruses can allow protein engineering, such as affi nity maturation of biomolecules. Hydrophobic and aggregation-prone, membrane proteins oft en prove too insoluble for conventional in vitro biochemical studies. To engineer soluble variants of human caveolin-1, a phage-displayed library of caveolin variants targeted the hydrophobic intra-membrane domain with substitutions to charged residues. Anti-selections for insolubility removed hydrophobic variants, and positive selections for binding to the known caveolin ligand HIV gp41 isolated functional, folded variants. Assays with several caveolin binding partners demonstrated the successful folding and functionality by a solubilized, full-length caveolin variant selected from the library. Th is caveolin variant allowed assay of the direct interaction between caveolin and cavin, an experiment requiring purifi ed proteins. Th e approach provides a general method for solubilization and engineering of membrane-associated proteins by phage display. In addition, the interaction between caveolin and HIV gp41 served as the starting point for investigating derivatives of the HIV fusion inhibitor peptide T20 (Fuzeon) as potential caveolin ligands. Phage-displayed wild-type T20 provided the scaff old for homolog shotgun scanning libraries for affi nity maturation of caveolin binders. Th e resulting variants have been shown to bind caveolin specifi cally and with high affi nity in both in vitro and cellular experiments.
Agnes Hajduczki has completed her undergraduate studies at the University of California, Los Angeles. She is currently working on her Ph.D. at the University of California, Irvine, in the Department of Molecular Biology and Biochemistry.