PARTIAL CHARACTERIZATION OF A HEPARAN SULFATE PROTEOGLYCAN RELEASED FROM PRECHONDROGENIC RAT LIMB BUDS BY HEPARIN

FUMIKO MATSUI, ATSUHIKO OOHIRA, RYUJIRO SHOJI
1991 Biomedical research  
Prechondrogenic rat limb buds contain two different molecular species of heparan sulfate proteoglycan, a smaller one (HSPG I»-A) and a larger one (HSPG III-A). Although a part of HSPG I-A was solubilized with phosphate-buffered saline (PBS) from the limb bud, HSPG III-A and a large part of HSPG I-A were resistant to extraction with the same solution. To examine whether these PBS-insoluble proteoglycans exist in a different way in the limb bud, proteoglycans were extracted from the PBS-insoluble
more » ... m the PBS-insoluble fraction of the limb bud, which had been labeled with [35S]sulfate, successively with the following solutions in the presence of protease inhibitors; heparin (100 ,ug/ml) in PBS, 0.8% nonanoyl-N-methyl-glucamide (MEGA-9) in PBS, and 4 M guanidine HCl in 50 mM Tris-HCl, pH 7.5. About 40% of [35S]labeled material in the PB S-insoluble fraction were solubilized with the heparin solution. About 25%, 30% and 5% were recovered in MEGA-9 extract, guanidine HCl extract, and the
doi:10.2220/biomedres.12.89 fatcat:6gjrll5umjc6horfmvfy3s2s3i