Data from Native adiponectin in serum binds to mammalian cells expressing T-cadherin, but not AdipoRs or calreticulin

Binding of APN Native APN binding study qPCR absolute quantification_2018-11-05 185355 (version 1).xlsx 2018-11-05 185355     2018-12-10 172652     2018-12-10 204623     human moouse serum HMW binding_2018-11-01 13h06m07s 10.400s Chemiluminescence   mAPN mSerum on mAPN recpetors 2018-10-30 13h43m23s 12.460s Chemiluminescence Figure 1C_mSerum to HEK OE   T-cad mSerum hSerum on APN-receptors 2018-10-30 13h29m33s 7.243s Chemiluminescence   T-cad mSerum hSerum on APN-receptors 2018-10-30 13h29m33s 7.243s Chemiluminescence Figure 1C_mSerum to HEK OE   Tubulin_mSerum on mAPN-receptors 293a 2018-10-30 13h24m42s 6.647s Chemiluminescence Figure 1C_mSerum to HEK OE   human moouse serum HMW binding_2018-11-01 13h06m07s 10.400s Chemiluminescence Figure 1C_mSerum to HEK OE     2018-10-30 13h24m42s 6.647s Chemiluminescence Figure 1C_mSerum to HEK OE raw data   2018-10-30 13h24m42s 6.647s Chemiluminescence+2018-10-30 13h25m23s 0.123s Colorimetric Figure 1C_mSerum to HEK OE raw data   data deposit for eLife All data files including raw data compressed. Pass word is 3737. Data deposit.7z Native adiponectin in serum binds to cells expressing T-cadherin, but not AdipoRs or calreticulin All raw and calculated data compressed. Data deposit for eLife.zip

Adiponectin is an adipocyte-derived atypically abundant circulating factor that protects various organs and tissues through its receptors, AdipoRs, calreticulin, and T-cadherin. To identify the major binding partner of circulating native adiponectin, we expressed these receptors on the surface of HEK293 cells. Adiponectin, either that in mouse or human serum, purified from serum, or produced by mammalian cells, bound to cells expressing T-cadherin, but not to those expressing AdipoR1 or calreticulin. The stable introduction of T-cadherin and AdipoR1 into CHO cells resulted in the cell surface localization of these receptors. Native adiponectin in serum bound to cells expressing T-cadherin, not to those expressing AdipoR1. The knockdown of T-cadherin, but not AdipoRs resulted in the significant attenuation of native adiponectin binding to C2C12 myotubes. Therefore, native adiponectin binding depended on the amount of T-cadherin expressed in HEK293 cells, CHO cells, and C2C12 myotubes. Collectively, our mammalian cell-based studies suggest that T-cadherin is the major binding partner of native adiponectin in serum.