C-Terminal Domain of the Human Zinc Transporter hZnT8 Is Structurally Indistinguishable from Its Disease Risk Variant (R325W)

• Published in: International journal of molecular sciences 2020-01, Vol.21 (3), p.926
• Main Authors: Ullah, Raheem, Shehzad, Aamir, Shah, Majid Ali, March, Matteo De, Ismat, Fouzia, Iqbal, Mazhar, Onesti, Silvia, Rahman, Moazur, McPherson, Michael J
• Format: Article
• Language: English
• Subjects: Beta cells; Binding; biophysical characterization; c-terminal domain; Chromatography; Cytoplasm; Diabetes; Diabetes mellitus; Diabetes mellitus (non-insulin dependent); Disease; disease risk variant; Domains; human zinc transporter hznt8; Insulin; Mutation; Pancreas; Polymorphism; Protein structure; Proteins; Risk; Secondary structure; Secretory vesicles; Tryptophan; Zinc; Zinc transporter
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21030926

The human zinc transporter 8 (hZnT8) plays important roles in the storage of insulin in the secretory vesicles of pancreatic β cells. hZnT8 consists of a transmembrane domain, with its N- and C-termini protruding into the cytoplasm. Interestingly, the exchange of arginine to tryptophan at position 325 in the C-terminal domain (CTD) increases the risk of developing type 2 diabetes mellitus (T2D). In the present study, the CTDs of hZnT8 (the wild-type (WT) and its disease risk variant (R325W)) were expressed, purified, and characterized in their native forms by biophysical techniques. The data reveal that the CTDs form tetramers which are stabilized by zinc binding, and exhibit negligible differences in their secondary structure content and zinc-binding affinities in solution. These findings provide the basis for conducting further structural studies aimed at unravelling the molecular mechanism underlying the increased susceptibility to develop T2D, which is modulated by the disease risk variant.