Glycation of PDGF results in decreased biological activity

Advanced glycation end products (AGEs) are formed by the non-enzymatic glycation of proteins by reducing carbohydrates or α-oxo-aldehydes such as glyoxal and methylglyoxal and further rearrangements, eliminations and oxidations. AGE-modifications alter peptide structure, function and stability and accumulate under several pathophysiological conditions such as diabetes and are considered a biomarker of ageing. PDGF is a major regulator of wound healing, which is impaired in hyperglycaemia and ageing. We analyzed whether glycated PDGF has impaired activity in cell culture models and occurs in human subjects. PDGF was AGE-modified by the α-oxo-aldehydes glyoxal and methylglyoxal, which was shown by Western-blotting using α-carboxymethyllysine (CML) or α-arginine-pyrimidine (Arg-Pyr) antibodies. In mouse AKR-2B fibroblasts, this AGE-modified PDGF exhibited reduced signalling to AKT and ERK resulting in decreased cell proliferation. In the human osteosarcoma cell line 143B, PDGF signalling towards the AKT-kinase was decreased when using modified PDGF-AA, -AB, and -BB whereas the constitutive active ERK was not affected. Secreted proteins from collagen-activated platelets from diabetic subjects contained more CML-modified proteins compared to healthy controls. PDGF protein as a platelet protein coprecipitated in immunoprecipitation experiments with α-CML-antiserum. In summary, our data suggest that AGE-modification of PDGF contributes to reduced wound healing in diabetic patients.