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Release of podocalyxin into the extracellular space. Role of metalloproteinases

AutorFernández, Darío ; Larrucea, Susana ; Nowakowski, Adam; Pericacho, Miguel ; Parrilla, Roberto L. ; Sánchez Ayuso, Matilde
Palabras claveHuman podocalyxin
Fate of podocalyxin-GFP
Extracellular podocalyxin
CHO cells
Tera-1 cells
Fecha de publicaciónago-2011
CitaciónBBA - Molecular Cell Research 1813(8):1504-1510(2011)
ResumenPodocalyxin (PODXL) is a type I membrane mucoprotein abundantly presented in the epithelial cells (podocytes) of kidney glomeruli where it plays an important role in maintaining the plasma filtration. PODXL is also expressed in other types of cells but its function is ignored. A recombinant soluble fragment of the PODXL ectodomain modifies the signaling of the membrane bound PODXL. Based on this antecedent, we aimed at investigating whether PODXL could be cleaved and released into the extracellular space as a soluble peptide. In this study, we used a fusion protein of human PODXL and green fluorescent protein expressed in CHO cells (CHO-PODXL-GFP) and a human tumor cell (Tera-1) inherently expressing PODXL. PODXL was detected by wide-field microscopy in the Golgi, the plasma membrane and in a vesicular form preferentially located at the leading edges of the cell and also progressing along the filopodium. We detected PODXL in the insoluble and soluble fractions of the extracellular medium of CHO-PODXL-GFP cells. Stimulation of protein kinase C (PKC) by Phorbol-12-myristate-13-acetate (PMA) enhanced the release of PODXL to the extracellular space whereas this effect was prevented either by inhibitors of PKC or specific inhibitors of matrix metalloproteinases. It is concluded that intact PODXL is released to the extracellular space as a cargo of microvesicles and also as a soluble cleaved fragment of ectodomain
Descripción7 páginas, 8 figuras, 3 figuras S, 1 tabla S -- PAGS nros. 1504-1510
Versión del editorhttp://dx.doi.org./10.1016/j.bbamcr.2011.05.009
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