Design of a PDZbody, a bivalent binder of the E6 protein from human papillomavirus

Scientific Reports
O Andreas KarlssonPer Jemth

Abstract

Chronic infection by high risk human papillomavirus (HPV) strains may lead to cancer. Expression of the two viral oncoproteins E6 and E7 is largely responsible for immortalization of infected cells. The HPV E6 is a small (approximately 150 residues) two domain protein that interacts with a number of cellular proteins including the ubiquitin ligase E6-associated protein (E6AP) and several PDZ-domain containing proteins. Our aim was to design a high-affinity binder for HPV E6 by linking two of its cellular targets. First, we improved the affinity of the second PDZ domain from SAP97 for the C-terminus of HPV E6 from the high-risk strain HPV18 using phage display. Second, we added a helix from E6AP to the N-terminus of the optimized PDZ variant, creating a chimeric bivalent binder, denoted PDZbody. Full-length HPV E6 proteins are difficult to express and purify. Nevertheless, we could measure the affinity of the PDZbody for E6 from another high-risk strain, HPV16 (Kd = 65 nM). Finally, the PDZbody was used to co-immunoprecipitate E6 protein from HPV18-immortalized HeLa cells, confirming the interaction between PDZbody and HPV18 E6 in a cellular context.

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Citations

Jan 19, 2018·Viruses·Irina Suarez, Gilles Trave
Jun 3, 2020·Medicinal Research Reviews·Avinash KumarSuvarna G Kini
Apr 22, 2020·Advanced Therapeutics·Nikolaj R ChristensenKristian Strømgaard
Feb 2, 2021·Frontiers in Molecular Biosciences·Simon Erlendsson, Kaare Teilum
Jun 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Lennox Chitsike, Penelope J Duerksen-Hughes
Jun 13, 2021·Protein Engineering, Design & Selection : PEDS·Chen T LiangWei Zhang

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Methods Mentioned

BETA
co-immunoprecipitation
NMR
phage display
isothermal titration calorimetry
transfection
transfections
protein assay

Software Mentioned

Origin

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