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Isolation and Characterization of Nanobodies against a Zinc-Transporting P-Type ATPase

1
Department of Biomedical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200 Copenhagen, Denmark
2
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark
3
Department of Experimental Medical Science, Lund University, Sölvegatan 19, SE-221 84 Lund, Sweden
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Antibodies 2018, 7(4), 39; https://doi.org/10.3390/antib7040039
Received: 17 October 2018 / Revised: 31 October 2018 / Accepted: 4 November 2018 / Published: 7 November 2018
(This article belongs to the Special Issue Nanobody)
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Abstract

P-type ATPases form a large and ubiquitous superfamily of ion and lipid transporters that use ATP (adenosine triphosphate) to carry out their function. The IB subclass (PIB-ATPases) allows flux of heavy metals and are key players in metal detoxification, critical for human health, crops, and survival of pathogens. Nevertheless, PIB-ATPases remain poorly understood at a molecular level. In this study, nanobodies (Nbs) are selected against the zinc-transporting PIB-ATPase ZntA from Shigella sonnei (SsZntA), aiming at developing tools to assist the characterization of the structure and function of this class of transporters. We identify six different Nbs that bind detergent stabilized SsZntA. We further assess the effect of the Nbs on the catalytic function of SsZntA, and find that five nanobodies associate without affecting the function, while one nanobody significantly reduces the ATPase activity. This study paves the way for more refined mechanistical and structural studies of zinc-transporting PIB-ATPases. View Full-Text
Keywords: P-type ATPase; nanobody; llama; Zinc-transport; Zinc-transporting P-ATPase; ZntA P-type ATPase; nanobody; llama; Zinc-transport; Zinc-transporting P-ATPase; ZntA
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Longhin, E.; Grønberg, C.; Hu, Q.; Duelli, A.S.; Andersen, K.R.; Laursen, N.S.; Gourdon, P. Isolation and Characterization of Nanobodies against a Zinc-Transporting P-Type ATPase. Antibodies 2018, 7, 39.

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