The Amino-terminal Domains of the Ezrin, Radixin, and Moesin (ERM) Proteins Bind Advanced Glycation End Products, an Interaction That May Play a Role in the Development of Diabetic Complications

Abstract

The presence of advanced glycation end products (AGEs) formed because of hyperglycemia in diabetic patients has been strongly linked to the development of diabetic complications and disturbances in cellular function. In this report, we describe the isolation and identification of novel AGE-binding proteins from diabetic rat kidneys. The proteins were purified by cation exchange and AGE-modified bovine serum albumin (AGE-BSA) affinity chromatography.

Advanced Glycation End Products Inhibit Tubulogenesis and Migration of Kidney Epithelial Cells in an Ezrin-Dependent Manner

“…We showed previously that LLC-PK1 cells, when treated with HGF in the presence of collagen I, form tubules between domes and that treatment with AGE-BSA inhibited tubulogenesis (8,9). This model has been reported previously to be ezrindependent (15).…”

“…Tubulogenesis experiments were performed as described previously (9). Briefly, LLC-PK1 cells were cultured in 24-well plates in 10% FCS/DMEM.…”

“…As with tubulogenesis, ezrin overexpression in LLC-PK1 cells enhances cell migration, whereas N-ezrin expression impairs HGF-induced migration (15). We recently showed that AGE-BSA inhibited LLC-PK1 tubulogenesis and ezrin tyrosine phosphorylation (9).…”

“…We recently described the novel binding of AGE to the ERM (ezrin, radixin, and moesin) family of proteins (9). ERM proteins are critical regulators of interactions between the cell membrane and the cytoskeleton (10).…”

Prohibitin-2 Binding Modulates Insulin-like Growth Factor-binding Protein-6 (IGFBP-6)-induced Rhabdomyosarcoma Cell Migration

“…Phosphorylation of N-ezrin (5 g) by the EGF receptor (1.62 units) was used as a positive control as described previously (16). The reaction was stopped by the addition of SDS-PAGE sample buffer and boiling, and the products were analyzed by SDS-10% PAGE and phosphotyrosine Western blotting.…”