Transmembrane transport of the Gαq protein carboxyl terminus imitation polypeptide GCIP-27

Abstract

The Gαq protein carboxyl terminus imitation polypeptide (GCIP)-27 has been shown to affect cardiac hypertrophy and vascular remodeling in various models both in vitro and in vivo. Transport across the plasma membrane is a critical step in regulating the action of this peptide drug. This study was designed to explore the mechanisms underlying the transmembrane transport of GCIP-27. The peptide drug was labeled with fluorescein isothiocyanate (FITC), and measured in a time- and concentration-dependent manner using laser confocal microscopy. Various transport inhibitors, including energy and endocytosis inhibitors, were used to identify the factors that regulate its transmembrane transport. GCIP-27 transport was examined in cardiomyocytes, cardiac fibroblasts, vascular endothelial cells, vascular smooth muscle cells (VSMCs) and hepatocytes. Atomic force microscopy and scanning electron microscopy were used to determine the ultrastructure of the cardiomyocyte membranes. The results showed that GCIP-27 was transported through the plasmalemma in a time- and concentration-dependent manner. The rate of uptake and the level of GCIP-27 in the cells decreased significantly after treatment with energy inhibitors, methyl-ß-cyclodextrin chlorpromazine or heparin. GCIP-27 levels in VSMCs and cardiomyocytes were significantly greater than the levels observed in hepatocytes, cardiac fibroblasts and vascular endothelial cells. Treatment with GCIP-27 led to a marked increase in the surface roughness of the cellular membrane. In conclusion, the transmembrane transport of GCIP-27 is mediated by endocytosis, which requires energy, and GCIP-27 preferentially enters myocardial cells and VSMCs.