Nanobodies as Probes for Protein Dynamics in Vitro and in Cells

Oleg Y. Dmitriev; Svetlana Lutsenko; Serge Muyldermans

doi:10.1074/jbc.R115.679811

Nanobodies as Probes for Protein Dynamics in Vitro and in Cells*

From the ^‡Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada,
the ^§Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, and
the ^¶Unit of Cellular and Molecular Immunology, Vrije Universiteit Brussel, 1050, Brussels, Belgium

↵1 To whom correspondence should be addressed: 107 Wiggins Rd., Saskatoon, Saskatchewan S7N 5E5, Canada. Tel.: 306-966-4377; E-mail: Oleg.Dmitriev{at}usask.ca.

Abstract

Nanobodies are the recombinant antigen-recognizing domains of the minimalistic heavy chain-only antibodies produced by camels and llamas. Nanobodies can be easily generated, effectively optimized, and variously derivatized with standard molecular biology protocols. These properties have triggered the recent explosion in the nanobody use in basic and clinical research. This review focuses on the emerging use of nanobodies for understanding and monitoring protein dynamics on the scales ranging from isolated protein domains to live cells, from nanoseconds to hours. The small size and high solubility make nanobodies uniquely suited for studying protein dynamics by NMR. The ability to produce conformation-sensitive nanobodies in cells enables studies that link structural dynamics of a target protein to its cellular behavior. The link between in vitro and in-cell dynamics, afforded by nanobodies, brings the analysis of such important events as receptor signaling, membrane protein trafficking, and protein interactions to the next level of resolution.

Footnotes

↵* This work was supported by the Canadian Institutes of Health Research and Saskatchewan Health Research Foundation grants (to O. Y. D.) and by National Institutes of Health Grant GM067166 (to S. L.). This is the second article in the Thematic Minireview series “Modern Technologies for In-cell Biochemistry.” The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the author and does not necessarily represent the official views of the National Institutes of Health.