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

doi:10.3390/antib7040039

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

Institut for Molekylærbiologi og Genetik - Plantemolekylærbiologi

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

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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

Originalsprog	Engelsk
Artikelnummer	39
Tidsskrift	Antibodies
Vol/bind	7
Nummer	4
Antal sider	15
ISSN	2073-4468
DOI	https://doi.org/10.3390/antib7040039
Status	Udgivet - 7 nov. 2018

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10.3390/antib7040039Licens: CC BY

antibodies-07-00039 (3)Forlagets udgivne version, 4,56 MBLicens: CC BY

Citationsformater

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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",

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T1 - Isolation and Characterization of Nanobodies against a Zinc-Transporting P-Type ATPase

AU - Longhin, Elena

AU - Grønberg, Christina

AU - Hu, Qiaoxia

AU - Duelli, Annette Susanne

AU - Andersen, Kasper Røjkjær

AU - Laursen, Nick Stub

AU - Gourdon, Pontus

N1 - This article belongs to the Special Issue Nanobody

PY - 2018/11/7

Y1 - 2018/11/7

N2 - 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

AB - 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

KW - P-type ATPase

KW - nanobody

KW - llama

KW - Zinc-transport

KW - Zinc-transporting P-ATPase

KW - ZntA

U2 - 10.3390/antib7040039

DO - 10.3390/antib7040039

M3 - Journal article

C2 - 31544889

SN - 2073-4468

VL - 7

JO - Antibodies

JF - Antibodies

IS - 4

M1 - 39

ER -

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

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