Structure-based design of a streptavidin mutant specific for an artificial biotin analogue

In this study, we improve the binding pocket of LISA-314 to abolish its affinity for endogenous BTN species, therefore ensuring that the newly designed LISA-314 binds only artificial BTN analogue. The replacement of three amino acid residues was performed in two steps to develop a mutant termed V212, which selectively binds to 6-(5-((3aS,4S,6aR)-2-iminohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexanoic acid (iminobiotin long tail, IMNtail). Surface plasmon resonance results showed that V212 has a Kd value of 5.9 x 10–7 M towards IMNtail, but no binding affinity for endogenous BTN species. This V212/IMNtail system will be useful as a novel delivery tool for anticancer therapy.

Source: Journal of Biochemistry - May 27, 2015 Category: Biochemistry Authors: Kawato, T., Mizohata, E., Shimizu, Y., Meshizuka, T., Yamamoto, T., Takasu, N., Matsuoka, M., Matsumura, H., Kodama, T., Kanai, M., Doi, H., Inoue, T., Sugiyama, A. Tags: Regular Papers Source Type: research