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

Elena Longhin, Christina Grønberg, Qiaoxia Hu, Annette Susanne Duelli, Kasper Røjkjær Andersen, Nick Stub Laursen, Pontus Gourdon

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Abstract

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
Original languageEnglish
Article number39
JournalAntibodies
Volume7
Issue4
Number of pages15
ISSN2073-4468
DOIs
Publication statusPublished - 7 Nov 2018

Keywords

  • P-type ATPase
  • nanobody
  • llama
  • Zinc-transport
  • Zinc-transporting P-ATPase
  • ZntA

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