Nesfatin-3 possesses divalent metal ion binding properties, which remain hidden in the nucleobindin-2 precursor protein

• Published in: Cell communication and signaling Vol. 21; no. 1; p. 165
• Main Authors: Skorupska-Stasiak, Anna, Bystranowska, Dominika, Tran, Józef Ba, Krężel, Artur, Ożyhar, Andrzej
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 29.06.2023; BioMed Central; BMC
• Subjects: Amino acids; Analysis; Comparative analysis; Food intake; Health aspects; Hypothalamus; Investigations; Ions; Ligands; Metal ion-induced conformational change; Metal ions; Mg2+/Ca2+Zn2+-binding proteins; Nesfatins; Nucb2; Nucleobindins; Peptides; Physiology; Protein binding; Proteins; Proteolytic processing of the precursor proteins; Zinc; Poland; Proteolytic processing of the precursor proteins; Nesfatins; Mg2+/Ca2+Zn2+-binding proteins; Nucb2; Metal ion-induced conformational change
• ISSN: 1478-811X; 1478-811X
• DOI: 10.1186/s12964-023-01181-6

Nucleobindin-2 (Nucb2) is a multidomain protein that, due to its structure, participates in many physiological processes. It was originally identified in several regions of the hypothalamus. However, more recent studies have redefined and extended the function of Nucb2 far beyond its initially observed role as a negative modulator of food intake. Previously, we described Nucb2 as structurally divided into two parts: the Zn -sensitive N-terminal half and the Ca -sensitive C-terminal half. Here, we investigated the structural and biochemical properties of its C-terminal half, which, after posttranslational processing, yields the formation of a fully uncharacterized peptide product known as nesfatin-3. Nesfatin-3 likely contains all the key respective structural regions of Nucb2. Hence, we expected that its molecular properties and affinity toward divalent metal ions might resemble those of Nucb2. Surprisingly, the obtained results showed that the molecular properties of nesftain-3 were completely different from those of its precursor protein. Moreover, we designed our work as a comparative analysis of two nesfatin-3 homologs. We noticed that in their apo forms, both proteins had similar shapes and existed in solution as extended molecules. They both interacted with divalent metal ions, and this interaction manifested itself in a compaction of the protein molecules. Despite their similarities, the differences between the homologous nesfatin-3s were even more informative. Each of them favored interaction with a different metal cation and displayed unique binding affinities compared either to each other or to Nucb2. The observed alterations suggested different from Nucb2 physiological roles of nesfatin-3 and different impacts on the functioning of the tissues and on metabolism and its control. Our results clearly demonstrated that nesfatin-3 possessed divalent metal ion binding properties, which remained hidden in the nucleobindin-2 precursor protein.