Crystal structure of streptavidin mutant with low immunogenicity

Tatsuya Kawato; Eiichi Mizohata; Tomohiro Meshizuka; Hirofumi Doi; Takeshi Kawamura; Hiroyoshi Matsumura; Kyohei Yumura; Kouhei Tsumoto; Tatsuhiko Kodama; Tsuyoshi Inoue; Akira Sugiyama

doi:10.1016/j.jbiosc.2014.10.025

Crystal structure of streptavidin mutant with low immunogenicity

J Biosci Bioeng. 2015 Jun;119(6):642-7. doi: 10.1016/j.jbiosc.2014.10.025. Epub 2014 Nov 26.

Affiliations

¹ Division of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
² Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo 153-8904, Japan.
³ Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan; Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa 277-8562, Japan.
⁴ Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan. Electronic address: sugiyama@lsbm.org.

PMID: 25434833
DOI: 10.1016/j.jbiosc.2014.10.025

Abstract

We previously created a low-immunogenic core streptavidin mutant No. 314 (LISA-314) by replacing six amino-acid residues for use as a delivery tool for an antibody multistep pre-targeting process (Yumura et al., Protein Sci., 22, 213-221, 2013). Here, we performed high-resolution X-ray structural analyses of LISA-314 and wild-type streptavidin to investigate the effect of substitutions on the protein function and the three-dimensional structure. LISA-314 forms a tetramer in the same manner as wild-type streptavidin. The binding mode of d-biotin in LISA-314 is also completely identical to that in wild-type streptavidin, and conformational changes were observed mostly at the side chains of substituted sites. Any large conformational changes corresponding to the reduction of B factors around the substituted sites were not observed. These results demonstrated the LISA-314 acquired low immunogenicity without losing structural properties of original wild-type streptavidin.

Keywords: B factor; Crystal structure; Low immunogenicity; Pre-targeting; Protein engineering; Streptavidin; X-ray structural analysis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Substitution / genetics
Biotin / chemistry
Biotin / immunology
Biotin / metabolism
Crystallography, X-Ray
Models, Molecular
Mutant Proteins / chemistry*
Mutant Proteins / genetics
Mutant Proteins / immunology*
Mutant Proteins / metabolism
Protein Binding
Protein Conformation
Protein Engineering
Streptavidin / chemistry*
Streptavidin / genetics
Streptavidin / immunology*
Streptavidin / metabolism

Substances

Mutant Proteins
Biotin
Streptavidin