RRC ID 65882
Author Fukuda S, Kita S, Obata Y, Fujishima Y, Nagao H, Masuda S, Tanaka Y, Nishizawa H, Funahashi T, Takagi J, Maeda N, Shimomura I.
Title The unique prodomain of T-cadherin plays a key role in adiponectin binding with the essential extracellular cadherin repeats 1 and 2.
Journal J Biol Chem
Abstract Adiponectin, an adipocyte-derived circulating protein, accumulates in the heart, vascular endothelium, and skeletal muscles through an interaction with T-cadherin (T-cad), a unique glycosylphosphatidylinositol-anchored cadherin. Recent studies have suggested that this interaction is essential for adiponectin-mediated cardiovascular protection. However, the precise protein-protein interaction between adiponectin and T-cad remains poorly characterized. Using ELISA-based and surface plasmon analyses, we report here that T-cad fused with IgG Fc as a fusion tag by replacing its glycosylphosphatidylinositol-anchor specifically bound both hexameric and larger multimeric adiponectin with a dissociation constant of ∼1.0 nm and without any contribution from other cellular or serum factors. The extracellular T-cad repeats 1 and 2 were critical for the observed adiponectin binding, which is required for classical cadherin-mediated cell-to-cell adhesion. Moreover, the 130-kDa prodomain-bearing T-cad, uniquely expressed on the cell surface among members of the cadherin family and predominantly increased by adiponectin, contributed significantly to adiponectin binding. Inhibition of prodomain-processing by a prohormone convertase inhibitor increased 130-kDa T-cad levels and also enhanced adiponectin binding to endothelial cells both by more preferential cell-surface localization and by higher adiponectin-binding affinity of 130-kDa T-cad relative to 100-kDa T-cad. The preferential cell-surface localization of 130-kDa T-cad relative to 100-kDa T-cad was also observed in normal mice aorta in vivo In conclusion, our study shows that a unique key feature of the T-cad prodomain is its involvement in binding of the T-cad repeats 1 and 2 to adiponectin and also demonstrates that adiponectin positively regulates T-cad abundance.
Volume 292(19)
Pages 7840-7849
Published 2017-5-12
DOI 10.1074/jbc.M117.780734
PII S0021-9258(20)41883-6
PMID 28325833
PMC PMC5427265
MeSH Adiponectin / chemistry* Adiponectin / genetics Animals CHO Cells Cadherins / chemistry* Calcium / chemistry Cell Adhesion Cell Membrane / metabolism Cricetinae Cricetulus Disulfides / chemistry Endothelial Cells / cytology Endothelial Cells / metabolism Enzyme-Linked Immunosorbent Assay Glycosylphosphatidylinositols / chemistry HEK293 Cells Humans Immunoglobulin G / chemistry Kinetics Male Mice Mice, Inbred C57BL Mice, Knockout Protein Binding Protein Domains Protein Interaction Mapping Surface Plasmon Resonance
IF 4.238
Human and Animal Cells 293(RCB1637)