SYNTHESIS OF PROTEOGLYCAN DURING HL-60 CELL-DIFFERENTIATION

Newly synthesized proteoglycans (PG) in proliferating HL-60 cells were labeled with [S-35]-sulfate and fractionated on DEAE-cellulose column s. Peak fractions were digested with chondroitinase ABC and separated by paper chromatography. Chondroitin sulfate was identified as the maj or PG. Treatment of cells with 16 nM TPA resulted in a 1.8-fold increa se in total sulfate incorporation into PG, a shift in its location fro m the cellular to the extracellular compartment, and an increase in th e total charge of PG (based on the profile of elution from DEAE-cellul ose columns), compared to controls. Incorporation of [H-3]-glucosamine into cellular PG was markedly decreased in TPA-treated cells; the sul fate/glucosamine ratio showed a 13.5-fold decrease in the newly synthe sized cellular PG. The sulfate/glucosamine ratio, however, was increas ed by 7.6-fold in secreted PG. (C) 1994 Academic Press, Inc.