E53, E96, D162, E247 and D322 in Ca<sup>2+</sup>-binding domains of annexin A2 are essential for regulating intracellular [Ca<sup>2+</sup>] and crystal adhesion to renal cells via ERK1/2 and JNK signaling pathways
3
Issued Date
2025-07-01
Resource Type
ISSN
00039861
eISSN
10960384
Scopus ID
2-s2.0-105001827774
Journal Title
Archives of Biochemistry and Biophysics
Volume
769
Rights Holder(s)
SCOPUS
Bibliographic Citation
Archives of Biochemistry and Biophysics Vol.769 (2025)
Suggested Citation
Yoodee S., Malaitad T., Plumworasawat S., Thongboonkerd V. E53, E96, D162, E247 and D322 in Ca<sup>2+</sup>-binding domains of annexin A2 are essential for regulating intracellular [Ca<sup>2+</sup>] and crystal adhesion to renal cells via ERK1/2 and JNK signaling pathways. Archives of Biochemistry and Biophysics Vol.769 (2025). doi:10.1016/j.abb.2025.110410 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/109491
Title
E53, E96, D162, E247 and D322 in Ca<sup>2+</sup>-binding domains of annexin A2 are essential for regulating intracellular [Ca<sup>2+</sup>] and crystal adhesion to renal cells via ERK1/2 and JNK signaling pathways
Author(s)
Author's Affiliation
Corresponding Author(s)
Other Contributor(s)
Abstract
Annexin A2 (ANXA2) is expressed inside the cytoplasm and on the surface of renal tubular epithelial cells (RTECs) and is documented as a calcium oxalate monohydrate (COM) crystal-binding protein. Nevertheless, its molecular mechanism involved in kidney stone disease (KSD) remains underinvestigated. Herein, we performed various molecular assays to unravel the roles of ANXA2 and core residues (E53, E96, D162, E247 and D322) in its Ca2+-binding domains in the stone formation mechanism, particularly at crystal-cell adhesion step and downstream signaling cascade. ANXA2 was up-regulated in apical membranes, not cytosol, of RTECs after COM crystal exposure. Neutralizing the surface expression of ANXA2 by a specific monoclonal antibody and silencing its expression by small interfering RNA (siRNA) significantly decreased COM crystal-cell adhesion. siRNA also suppressed the COM-induced up-regulation of phospho-ERK1/2 and phospho-JNK, but not that of phospho-p38. Overexpression of ANXA2 wild-type (WT), but not that of E53A, E96A, D162A, E247A and D322A mutants of its Ca2+-binding domains, significantly increased intracellular [Ca2+], COM-cell adhesion, and phospho-ERK1/2 level. Therefore, E53, E96, D162, E247 and D322 in the Ca2+-binding domains of annexin A2 are essential for regulating intracellular [Ca2+] and COM crystal-cell adhesion via ERK1/2 and JNK signaling pathways.