TY - JOUR
T1 - Twigged streptavidin polymer as a scaffold for protein assembly
AU - Matsumoto, Takuya
AU - Isogawa, Yuki
AU - Minamihata, Kosuke
AU - Tanaka, Tsutomu
AU - Kondo, Akihiko
N1 - Funding Information:
This work was supported in part by a Grant-in-Aid for Young Scientists B ( 15K18276 ) from the Japan Society for the Promotion of Science (JSPS), and by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe), MEXT, Japan.
Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2016/5/10
Y1 - 2016/5/10
N2 - Protein assemblies are an emerging tool that is finding many biological and bioengineering applications. We here propose a method for the site-specific assembly of proteins on a twigged streptavidin (SA) polymer using streptavidin as a functional scaffold. SA was genetically appended with a G tag (sortase A recognition sequence) and a Y tag (HRP recognition sequence) on its N- and C-termini, respectively, to provide G-SA-Y. G-SA-Y was polymerized using HPR-mediated tyrosine coupling, then fluorescent proteins were immobilized on the polymer by biotin-SA affinity and sortase A-mediated ligation. Fluorescence measurements showed that the proteins were immobilized in close proximity to each other. Hydrolyzing enzymes were also functionally assembled on the G-SA-Y polymer. The site-specific assembly of proteins on twigged SA polymer may find new applications in various biological and bioengineering fields.
AB - Protein assemblies are an emerging tool that is finding many biological and bioengineering applications. We here propose a method for the site-specific assembly of proteins on a twigged streptavidin (SA) polymer using streptavidin as a functional scaffold. SA was genetically appended with a G tag (sortase A recognition sequence) and a Y tag (HRP recognition sequence) on its N- and C-termini, respectively, to provide G-SA-Y. G-SA-Y was polymerized using HPR-mediated tyrosine coupling, then fluorescent proteins were immobilized on the polymer by biotin-SA affinity and sortase A-mediated ligation. Fluorescence measurements showed that the proteins were immobilized in close proximity to each other. Hydrolyzing enzymes were also functionally assembled on the G-SA-Y polymer. The site-specific assembly of proteins on twigged SA polymer may find new applications in various biological and bioengineering fields.
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U2 - 10.1016/j.jbiotec.2016.03.030
DO - 10.1016/j.jbiotec.2016.03.030
M3 - Article
C2 - 27002233
AN - SCOPUS:84962284814
SN - 0168-1656
VL - 225
SP - 61
EP - 66
JO - Journal of Biotechnology
JF - Journal of Biotechnology
ER -