TY - GEN
T1 - Site-selective immobilization of streptavidin on enzymatically biotinylated bacterial magnetic particles
AU - Maeda, Yoshiaki
AU - Yoshino, Tomoko
AU - Takeyama, Haruko
AU - Takahashi, Masaaki
AU - Ginya, Harumi
AU - Asahina, Junko
AU - Tajima, Hideji
AU - Matsunaga, Tadashi
PY - 2008/12/1
Y1 - 2008/12/1
N2 - Biotinylated magnetic nanoparticles were constructed by displaying biotin acceptor peptide (BAP) on the surfaces of bacterial magnetic particles (BacMPs) synthesized by Magnetospirillum niagneticum AMB-1. Both BAP and green fluorescent protein (GFP) were fused to Mmsl3 that was isolated from BacMP membranes. The localization of the fusion protein, BAP-Mmsl3-GFP, was confirmed by fluorescence analysis. BacMPs that expressed B AP-Mms 13-GFP (BAP/GFP-BacMPs) were extracted from bacterial cells and incubated with biotin and Escherichia coli biotin ligase. The in vitro biotinylation of BAP/GFP-BacMPs was confirmed using alkaline phosphatase (ALP)-labeled streptavidin. The conjugation system developed in this study provides a method for producing biotin- or streptavidin-labeled magnetic nanoparticles without the use of a crosslinker reagent. Various functional materials can be immobilized site-selectively onto these uniquely designed BacMPs. By combining this site-selective biotinylation technology and protein display methodology, increasingly innovative and attractive magnetic nano-materials can be constructed.
AB - Biotinylated magnetic nanoparticles were constructed by displaying biotin acceptor peptide (BAP) on the surfaces of bacterial magnetic particles (BacMPs) synthesized by Magnetospirillum niagneticum AMB-1. Both BAP and green fluorescent protein (GFP) were fused to Mmsl3 that was isolated from BacMP membranes. The localization of the fusion protein, BAP-Mmsl3-GFP, was confirmed by fluorescence analysis. BacMPs that expressed B AP-Mms 13-GFP (BAP/GFP-BacMPs) were extracted from bacterial cells and incubated with biotin and Escherichia coli biotin ligase. The in vitro biotinylation of BAP/GFP-BacMPs was confirmed using alkaline phosphatase (ALP)-labeled streptavidin. The conjugation system developed in this study provides a method for producing biotin- or streptavidin-labeled magnetic nanoparticles without the use of a crosslinker reagent. Various functional materials can be immobilized site-selectively onto these uniquely designed BacMPs. By combining this site-selective biotinylation technology and protein display methodology, increasingly innovative and attractive magnetic nano-materials can be constructed.
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M3 - Conference contribution
AN - SCOPUS:77957940490
SN - 9781605608488
T3 - Materials Research Society Symposium Proceedings
SP - 65
EP - 70
BT - From Biological Materials to Biomimetic Material Synthesis
T2 - 2008 MRS Spring Meeting
Y2 - 24 March 2008 through 28 March 2008
ER -