4ATZ

Ad5 knob in complex with a designed ankyrin repeat protein

Summary for 4ATZ

• Entry DOI: 10.2210/pdb4atz/pdb
• Related: 1KNB
• Descriptor: Fiber protein, DESIGNED ANKYRIN REPEAT PROTEIN (3 entities in total)
• Functional Keywords: viral protein-de novo protein complex, darpin, protein design, viral protein/de novo protein
• Biological source: Human adenovirus C serotype 5 (HAdV-5); More
• Total number of polymer chains: 6
• Total formula weight: 115085.82

Authors

Mittl, P.R.E.,Hess, C.,Dreier, B. (deposition date: 2012-05-11, release date: 2013-02-27, Last modification date: 2023-12-20)

Primary citation

Dreier, B.,Honegger, A.,Hess, C.,Nagy-Davidescu, G.,Mittl, P.R.,Grutter, M.G.,Belousova, N.,Mikheeva, G.,Krasnykh, V.,Pluckthun, A.
Development of a generic adenovirus delivery system based on structure-guided design of bispecific trimeric DARPin adapters.
Proc. Natl. Acad. Sci. U.S.A., 110:E869-E877, 2013

PubMed Abstract: Adenoviruses (Ads) have shown promise as vectors for gene delivery in clinical trials. Efficient viral targeting to a tissue of choice requires both ablation of the virus' original tropism and engineering of an efficient receptor-mediated uptake by a specific cell population. We have developed a series of adapters binding to the virus with such high affinity that they remain fully bound for >10 d, block its natural receptor binding site and mediate interaction with a surface receptor of choice. The adapter contains two fused modules, both consisting of designed ankyrin repeat proteins (DARPins), one binding to the fiber knob of adenovirus serotype 5 and the other binding to various tumor markers. By solving the crystal structure of the complex of the trimeric knob with three bound DARPins at 1.95-Å resolution, we could use computer modeling to design a link to a trimeric protein of extraordinary kinetic stability, the capsid protein SHP from the lambdoid phage 21. We arrived at a module which binds the knob like a trimeric clamp. When this clamp was fused with DARPins of varying specificities, it enabled adenovirus serotype 5-mediated delivery of a transgene in a human epidermal growth factor receptor 2-, epidermal growth factor receptor-, or epithelial cell adhesion molecule-dependent manner with transduction efficiencies comparable to or even exceeding those of Ad itself. With these adapters, efficiently produced in Escherichia coli, Ad can be converted rapidly to new receptor specificities using any ligand as the receptor-binding moiety. Prefabricated Ads with different payloads thus can be retargeted readily to many cell types of choice.

PubMed: 23431166
DOI: 10.1073/pnas.1213653110

Experimental method

X-RAY DIFFRACTION (1.95 Å)