Differential transport kinetics of chondroitin sulfate and dermatan sulfate proteoglycan by monkey aorta smooth muscle cells

Abstract

Pulse-chase studies were performed to study the kinetics of chondroitin sulfate proteoglycan (CSPG) and dermatan sulfate proteoglycan (DSPG) transport in monkey aorta smooth muscle cells. During a short pulse (5 min) with [ 35S]Na 2SO 4 (500 μCi/ml), the cells synthesized 59% DSPG, 38% CSPG, and 3% heparan sulfate proteoglycan. Both DSPG and CSPG were transported out of the cell very rapidly after sulfate incorporation. At various chase times, proteoglycans (PGs) were isolated from four cellular compartments: (a) medium, (b) total cell lysate, (c) intracellular pool, and (d) extracellular pool. The PGs from the different pools were analyzed by Sepharose CL-2B column chromatography. The data of intracellular DSPG loss fitted a double exponential decay model: approximately 90% was secreted quickly with a t 1 2 of 7 min, and the remaining 10% had a dramatically slower rate of secretion ( t 1 2 of 130 min). DSPG was rapidly secreted into the medium without prior accumulation in the extracellular matrix. In contrast, the loss of intracellular CSPG fitted a single exponential decay model with a t 1 2 of 8 min; however, there was a significant accumulation of CSPG in the extracellular matrix compartment before release into the medium, resulting in a relatively slower secretion of CSPG into the medium ( t 1 2 of about 31 min). This delay in CSPG secretion into the medium is probably due to aggregation in the extracellular matrix, since addition of short hyaluronan oligomers (8–14 oligosaccharides) to the medium during the chase increased the rate of CSPG being secreted into the medium. We concluded that in aortic smooth muscle cell cultures, CSPG and DSPG are secreted via two distinct pathways through the cellular compartments.