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Isolation and Characterization of Nanobodies against a Zinc-Transporting P-Type ATPase.
Longhin, Elena; Grønberg, Christina; Hu, Qiaoxia; Duelli, Annette Susanne; Andersen, Kasper Røjkjær; Laursen, Nick Stub; Gourdon, Pontus.
Afiliación
  • Longhin E; Department of Biomedical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200 Copenhagen, Denmark. elonghin@sund.ku.dk.
  • Grønberg C; Department of Biomedical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200 Copenhagen, Denmark. christina.groenberg@sund.ku.dk.
  • Hu Q; Department of Biomedical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200 Copenhagen, Denmark. qiaoxia@sund.ku.dk.
  • Duelli AS; Department of Biomedical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200 Copenhagen, Denmark. duelli@sund.ku.dk.
  • Andersen KR; Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark. kra@mbg.au.dk.
  • Laursen NS; Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark. nsl@mbg.au.dk.
  • Gourdon P; Department of Biomedical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200 Copenhagen, Denmark. pontus@sund.ku.dk.
Antibodies (Basel) ; 7(4)2018 Nov 07.
Article en En | MEDLINE | ID: mdl-31544889
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Antibodies (Basel) Año: 2018 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Suiza
Texto completo
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XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Antibodies (Basel) Año: 2018 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Suiza