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

被引：6

作者：

Longhin, Elena ^{[1
]}

Gronberg, Christina ^{[1
]}

Hu, Qiaoxia ^{[1
]}

Duelli, Annette Susanne ^{[1
]}

Andersen, Kasper Rojkjaer ^{[2
]}

Laursen, Nick Stub ^{[2
]}

Gourdon, Pontus ^{[1
,3
]}

机构：

[1] Univ Copenhagen, Dept Biomed Sci, Blegdamsvej 3B, DK-2200 Copenhagen, Denmark

[2] Aarhus Univ, Dept Mol Biol & Genet, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark

[3] Lund Univ, Dept Expt Med Sci, Solvegatan 19, SE-22184 Lund, Sweden

来源：

ANTIBODIES | 2018年 / 7卷 / 04期

基金：

瑞典研究理事会;

关键词：

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

D O I：

10.3390/antib7040039

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

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 (P-IB-ATPases) allows flux of heavy metals and are key players in metal detoxification, critical for human health, crops, and survival of pathogens. Nevertheless, P-IB-ATPases remain poorly understood at a molecular level. In this study, nanobodies (Nbs) are selected against the zinc-transporting P-IB-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 P-IB-ATPases.

引用

页数：15

共 26 条

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