Evaluation of the roles of the cytosolic N-terminus and His-rich loop of ZNT proteins using ZNT2 and ZNT3 chimeric mutants

Scientific Reports
Kazuhisa FukueTaiho Kambe

Abstract

The physiological roles of Zn transporter (ZNT) proteins are being increasingly recognized, and three dimensional structures of ZNT bacterial homologs have facilitated our understanding of their biochemical characteristics at the molecular level. However, the biological role of the unique structural features of vertebrate ZNTs, which are absent in their bacterial homologues, is not completely understood. These ZNT sequences include a cytosolic His-rich loop between transmembrane helices IV and V and the cytosolic N-terminus. This study investigated the contribution of these features to zinc transport by ZNT proteins. The importance of the His residues in the cytosolic His-rich loop was investigated using ZNT2 Ala substitution and deletion mutants. The presence of His residues was not essential for zinc transport, even though they possibly participate in modulation of zinc transport activity. Furthermore, we determined the role of the N-terminus by characterizing ZNT2 and ZNT3 domain-swapped and deletion mutants. Unexpectedly, the N-terminus was also not essential for zinc transport by ZNT2 and the domain-swapped ZNT2 mutant, in which the cytosolic His-rich loop was substituted with that of ZNT3. These results provide molecular ...Continue Reading

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Citations

Mar 18, 2020·The Journal of Biological Chemistry·Eisuke SuzukiTaiho Kambe
Sep 1, 2019·The Journal of Biological Chemistry·Yukina Nishito, Taiho Kambe
Apr 17, 2019·Acta Crystallographica. Section D, Structural Biology·Camila A CotrimDavid Drew
Feb 1, 2021·The Journal of Biological Chemistry·Eisuke SuzukiTaiho Kambe
Mar 6, 2021·Molecular and Cellular Biochemistry·Carla P Barragán-ÁlvarezNestor E Díaz-Martínez

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Methods Mentioned

BETA
X-ray

Software Mentioned

ImageQuant
BLASTP
ClustalW

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