3WZO

Crystal structure of the core streptavidin mutant V21 (Y22S/N23D/S27D/Y83S/R84K/E101D/R103K/E116N) complexed with biotin long tail (BTNtail) at 1.5 A resolution

Summary for 3WZO

• Entry DOI: 10.2210/pdb3wzo/pdb
• Related: 3WZN 3WZP 3WZQ
• Descriptor: Streptavidin, 6-({5-[(3aS,4S,5S,6aR)-5-oxido-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanoyl}amino)hexanoic acid, GLYCEROL, ... (6 entities in total)
• Functional Keywords: beta-barrel, biotin binding protein
• Biological source: Streptomyces avidinii
• Cellular location: Secreted: P22629
• Total number of polymer chains: 4
• Total formula weight: 56255.47

Authors

Kawato, T.,Mizohata, E.,Shimizu, Y.,Meshizuka, T.,Yamamoto, T.,Takasu, N.,Matsuoka, M.,Matsumura, H.,Tsumoto, K.,Kodama, T.,Kanai, M.,Doi, H.,Inoue, T.,Sugiyama, A. (deposition date: 2014-10-01, release date: 2015-02-18, Last modification date: 2023-11-08)

Primary citation

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.
Structure-based design of a streptavidin mutant specific for an artificial biotin analogue.
J.Biochem., 157:467-475, 2015

PubMed Abstract: For a multistep pre-targeting method using antibodies, a streptavidin mutant with low immunogenicity, termed low immunogenic streptavidin mutant No. 314 (LISA-314), was produced previously as a drug delivery tool. However, endogenous biotins (BTNs) with high affinity (Kd < 10(-10) M) for the binding pocket of LISA-314 prevents access of exogenous BTN-labelled anticancer drugs. 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 × 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.

PubMed: 25645976
DOI: 10.1093/jb/mvv004

Experimental method

X-RAY DIFFRACTION (1.5 Å)