Nanobody-Displaying Flagellar Nanotubes

Scientific Reports
Ágnes KleinFerenc Vonderviszt

Abstract

In this work we addressed the problem how to fabricate self-assembling tubular nanostructures displaying target recognition functionalities. Bacterial flagellar filaments, composed of thousands of flagellin subunits, were used as scaffolds to display single-domain antibodies (nanobodies) on their surface. As a representative example, an anti-GFP nanobody was successfully inserted into the middle part of flagellin replacing the hypervariable surface-exposed D3 domain. A novel procedure was developed to select appropriate linkers required for functional internal insertion. Linkers of various lengths and conformational properties were chosen from a linker database and they were randomly attached to both ends of an anti-GFP nanobody to facilitate insertion. Functional fusion constructs capable of forming filaments on the surface of flagellin-deficient host cells were selected by magnetic microparticles covered by target GFP molecules and appropriate linkers were identified. TEM studies revealed that short filaments of 2-900 nm were formed on the cell surface. ITC and fluorescent measurements demonstrated that the fusion protein exhibited high binding affinity towards GFP. Our approach allows the development of functionalized flagel...Continue Reading

References

Sep 5, 1976·Journal of Molecular Biology·R Kamiya, S Asakura
Oct 20, 1991·Journal of Molecular Biology·F VondervisztK Namba
Sep 5, 1989·Journal of Molecular Biology·F VondervisztK Namba
Jan 1, 1974·Journal of Supramolecular Structure·T Iino
Jan 1, 1994·International Reviews of Immunology·B A Stocker, S M Newton
Dec 24, 1996·Proceedings of the National Academy of Sciences of the United States of America·Y Mimori-KiyosueK Namba
Jan 23, 2003·Protein Engineering·Richard A George, Jaap Heringa
Dec 22, 2005·Nature Biotechnology·Jean-Denis PédelacqGeoffrey S Waldo
Aug 1, 2007·Journal of Nanoscience and Nanotechnology·Mudalige Thilak KumaraBrian C Tripp
Apr 4, 2008·Proteins·Krishnakumar M Ravikumar, Wonmuk Hwang
Aug 12, 2008·Current Opinion in Pharmacology·Dirk SaerensSerge Muyldermans
Nov 26, 2008·Veterinary Immunology and Immunopathology·S MuyldermansD Saerens
Jun 17, 2009·Medical Microbiology and Immunology·Janusz WesolowskiFriedrich Koch-Nolte
Dec 17, 2009·Nature Structural & Molecular Biology·Axel KirchhoferUlrich Rothbauer
Sep 28, 2010·Journal of Molecular Biology·Adél MuskotálFerenc Vonderviszt
Oct 15, 2010·Protein Science : a Publication of the Protein Society·Marta H KubalaBrett M Collins
Feb 4, 2012·Protein Engineering, Design & Selection : PEDS·Agnes KleinFerenc Vonderviszt
Nov 13, 2012·PloS One·Olesya V StepanenkoKonstantin K Turoverov
Jan 2, 2013·Frontiers in Bioscience (Elite Edition)·Jay Russell MeredithRavi Aggarwal
Aug 2, 2013·Journal of Nanoscience and Nanotechnology·Piyush Kar, Karthik Shankar
Sep 26, 2013·Biochimica Et Biophysica Acta·Tohru Minamino
May 15, 2015·Molecular Biotechnology·Ágnes KleinFerenc Vonderviszt
Jul 20, 2016·EBioMedicine·Isabel Van Audenhove, Jan Gettemans
Aug 9, 2016·International Journal of Nanomedicine·Yongzhong WangLi Yu
Nov 3, 2016·Chembiochem : a European Journal of Chemical Biology·Éva Bereczk-TompaFerenc Vonderviszt

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Citations

Nov 23, 2019·Angewandte Chemie·Benedikt JedlitzkeHenning D Mootz
Oct 16, 2019·Proceedings of the National Academy of Sciences of the United States of America·Maniraj BhagawatiHenning D Mootz
Sep 2, 2020·The Journal of Biological Chemistry·Ross W ChelohaHidde L Ploegh
Feb 12, 2021·Scientific Reports·Hajnalka JankovicsFerenc Vonderviszt
Sep 28, 2020·Biochimica Et Biophysica Acta. General Subjects·Troy BurtchettBrian C Tripp

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Methods Mentioned

BETA
PCR
electrophoresis
transmission electron microscopy
electron microscopy
optical microscopy

Software Mentioned

MicroCal Origin

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