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Hyperglycemia decreased APT1 activity in HUVECs, due in part to altered acetylation of the APT1 protein. Click chemistry analyses demonstrated increased R-Ras palmitoylation in the setting of hyperglycemia. Altered R-Ras trafficking, increased R-Ras palmitoylation, and fibronectin retention were found in diabetes models. Loss of R-Ras depalmitoylation caused by APT1 deficiency constrained R-Ras membrane trafficking as shown by TIRF imaging. To rescue cellular phenotypes, we generated an R-Ras molecule with an inserted hydrophilic domai