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DERMATAN SULFATE IS THE MAJOR GLYCOSAMINOGLYCAN SYNTHESIZED BY CULTURED GUINEA PIG SKIN FIBROBLASTS
Author(s) -
Nakamura Toshitaka,
Shinkai Hiroshi
Publication year - 1985
Publication title -
the journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.9
H-Index - 65
eISSN - 1346-8138
pISSN - 0385-2407
DOI - 10.1111/j.1346-8138.1985.tb02880.x
Subject(s) - dermatan sulfate , glycosaminoglycan , hyaluronic acid , glucosamine , chemistry , biochemistry , chondroitin , hyaluronidase , chondroitin sulfate , heparan sulfate , guinea pig , sulfation , cell culture , biology , enzyme , anatomy , genetics , endocrinology
Fibroblasts from guinea pig skin were incubated with [ 3 H] glucosamine. The cells secreted [ 3 H] galactosamine‐labeled compounds predominantly as dermatan sulfate, which accounted for 65–75% of the total glycosaminoglycans. The majority production of dermatan sulfate was observed regardless of the composition of the culture medium, cell growing phase or age of animal. The labeled glycosaminoglycans were identified by enzymatic (chondroitinase ABC, chondroitinase AC and hyaluronidase) or nitrous acid treatment, followed by fluorography of two‐dimensional electrophoresis on cellulose acetate membranes. The molecular weight of the newly synthesized dermatan sulfate from medium of a confluent stage was calculated as 15,000 to 17,500 from molecular sieve chromatography. The radioactive dermatan sulfate was treated with chondroitinase ABC, yielding 68.4% Δ Di‐4S, 16.2% Δ Di‐6S, 13.3% Δ Di‐OS and 2% di‐sulfated hexuronosyl‐N‐acetylhexosamine units. Chondroitin sulfate was detected on a cellulose acetate membrane as a minor component in the culture medium from the cell growing stage. It was not detected in medium from a confluent stage. Organ culture of guinea pig skin or cell culture of human skin fibroblasts incorporated [ 3 H] glucosamine mainly into hyaluronic acid. About 90% of newly synthesized glycosaminoglycans was accounted for by hyaluronate.