In vivo biotinylation of bacterial magnetic particles by a truncated form of Escherichia coli biotin ligase and biotin acceptor peptide.

Applied and Environmental Microbiology
Yoshiaki MaedaTadashi Matsunaga

Abstract

Escherichia coli biotin ligase can attach biotin molecules to a lysine residue of biotin acceptor peptide (BAP), and biotinylation of particular BAP-fused proteins in cells was carried out by coexpression of E. coli biotin ligase (in vivo biotinylation). This in vivo biotinylation technology has been applied for protein purification, analysis of protein localization, and protein-protein interaction in eukaryotic cells, while such studies have not been reported in bacterial cells. In this study, in vivo biotinylation of bacterial magnetic particles (BacMPs) synthesized by Magnetospirillum magneticum AMB-1 was attempted by heterologous expression of E. coli biotin ligase. To biotinylate BacMPs in vivo, BAP was fused to a BacMP surface protein, Mms13, and E. coli biotin ligase was simultaneously expressed in the truncated form lacking the DNA-binding domain. This truncation-based approach permitted the growth of AMB-1 transformants when biotin ligase was heterologously expressed. In vivo biotinylation of BAP on BacMPs was confirmed using an alkaline phosphatase-conjugated antibiotin antibody. The biotinylated BAP-displaying BacMPs were then exposed to streptavidin by simple mixing. The streptavidin-binding capacity of BacMPs bioti...Continue Reading

References

Oct 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·K P WilsonB W Matthews
Jan 1, 1990·Methods in Enzymology·N M Green
Mar 15, 1981·Journal of Molecular Biology·D F Barker, A M Campbell
Apr 22, 1999·Protein Science : a Publication of the Protein Society·D BeckettP J Schatz
Nov 21, 2001·Protein Science : a Publication of the Protein Society·A Chapman-SmithJ C Wallace
Jun 13, 2003·Proceedings of the National Academy of Sciences of the United States of America·Ernie de BoerJohn Strouboulis
Jul 4, 2003·Applied and Environmental Microbiology·Yoshiko OkamuraTadashi Matsunaga
Jun 4, 1962·Biochemical and Biophysical Research Communications·D P KOSOW, M D LANE
May 7, 2004·Applied and Environmental Microbiology·Tomoko YoshinoTadashi Matsunaga
Dec 18, 2004·Science·Shiping Liao, Nadrian C Seeman
May 18, 2005·Proceedings of the National Academy of Sciences of the United States of America·Mark HowarthAlice Y Ting
Nov 18, 2005·Biochemical and Biophysical Research Communications·Tomoko Yoshino, Tadashi Matsunaga
Jan 5, 2006·Applied and Environmental Microbiology·Tomoko Yoshino, Tadashi Matsunaga
Nov 10, 2006·Nature·Jianlong WangStuart H Orkin
Jun 19, 2008·Journal of the Royal Society, Interface·Atsushi ArakakiTadashi Matsunaga
Jun 24, 2008·Applied and Environmental Microbiology·Yoshiaki MaedaTadashi Matsunaga
Jun 25, 2008·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Antoine ViensVasily Ogryzko
Jun 28, 2008·Journal of the American Chemical Society·Marta Fernández-SuárezAlice Y Ting
Dec 8, 2009·Nucleic Acids Research·Siew Loon OoiSteven Henikoff

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Citations

Jul 19, 2011·Applied and Environmental Microbiology·Anna PollithyDirk Schüler
Jan 30, 2014·Biochemical and Biophysical Research Communications·Yasuhiro SugamataTomoko Yoshino
Nov 6, 2013·The Journal of Biological Chemistry·Vandana Chakravartty, John E Cronan

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