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Updated: Oct 2, 2026

RhoC GTPase Activation Assay
Published on: August 22, 2010
Involvement of the GTPase Rho in the cellular uptake of low density lipoprotein by human skin fibroblasts
N Hrboticky1, T Feldmeer, M Essler
1Institut für Prophylaxe und Epidemiologie der Kreislaufkrankheiten, Universität München, Munich, Germany.
Abstract:
Members of the Rho subfamily of small GTPases have been implicated in the regulation of endocytosis of ligand/receptor complexes localised to clathrin-coated pits. In this paper, we investigated the role of Rho A in the post-receptor regulation of cellular uptake and metabolism of native low density lipoprotein (LDL) by primary human skin fibroblasts. Incubations of cells with the selective Rho GTPase inhibitor C3-transferase (C3T) upregulated the binding, lysosomal degradation and cell accumulation of labelled LDL. The rate of internalisation of surface-bound LDL was also higher in C3T-treated cells. Single cell injections with C3T and dominant active V14Rho confirmed the negative regulation of LDL uptake by Rho. While cells injected with C3T internalised more 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (diI)-labelled LDL, diI-LDL internalisation was dramatically suppressed in cells injected with the constitutively active V14Rho. The negative regulation of LDL uptake by Rho appeared to be independent of changes in the actin cytoskeleton. An increasing number of naturally occurring toxins and serum factors have been shown to influence Rho GTPase signalling cascades. The herein described post-translational regulation of LDL internalisation may modulate cell events occurring subsequent to cellular lipoprotein uptake.
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