Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells.

The serglycin proteoglycan is best known as a hematopoietic cell granule proteoglycan. It has been found that serglycin is synthesized by endothelial cells, is localized to cytoplasmic vesicles, and is constitutively secreted. Serglycin messenger RNA in human umbilical vein endothelial cells (HUVECs) and cultured human aortic endothelial cells was detected by reverse transcription-polymerase chain reaction. (35)S-sulfate-labeled secreted and intracellular proteoglycans were analyzed. It was found that 85% of the proteoglycans synthesized during culture were secreted. A core protein of the appropriate size for serglycin was detected by analysis of the chondroitinase-digested (35)S-sulfate-labeled HUVEC proteoglycans. This was the major core protein of the secreted chondroitin sulfate proteoglycans. Recombinant serglycin core protein was used to generate an antibody in chickens. A core protein identified by Western blotting of chondroitinase digests of HUVEC proteoglycans corresponded to the major (35)S-sulfate- labeled core protein. Identical results were obtained with 2 hematopoietic cell lines. Cyto-immunofluorescence showed cytoplasmic vesicular and perinuclear labeling in hematopoietic cells and HUVECs. The serglycin-containing vesicles in HUVECs are distinct from the Weibel-Palade bodies, which contain von Willebrand factor. Confocal microscopy showed that tissue plasminogen activator was distributed similarly to serglycin. Serglycin may be important for the function of these vesicles and, once secreted, for the modulation of the activity of their constituents.