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

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    Elena Longhin, Department of Biomedical Sciences, University of Copenhagen, Christina Grønberg, Department of Biomedical Sciences, University of Copenhagen, UkendtQiaoxia Hu, Department of Biomedical Sciences, University of Copenhagen, Annette Susanne Duelli, Department of Biomedical Sciences, University of Copenhagen,
  • Kasper Røjkjær Andersen
  • Nick Stub Laursen
  • Pontus Gourdon, Department of Experimental Medical Science, Lund University, Department of Biomedical Sciences, University of Copenhagen
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
OriginalsprogEngelsk
Artikelnummer39
TidsskriftAntibodies
Vol/bind7
Tidsskriftsnummer4
Antal sider15
ISSN2073-4468
DOI
StatusUdgivet - 7 nov. 2018

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