TY - JOUR
T1 - Extracellular matrix (ECM) proteoglycans produced by cultured bovine corneal endothelial cells
AU - Nevo, Zvi
AU - Gonzalez, Robert
AU - Gospodarowicz, Denis
N1 - Funding Information:
This work was supported by Grant EY 02186 from the National Institutes of Health. The authors wish to thank Mr. Harvey Scodel for his invaluable help in the preparation of this manuscript.
PY - 1984
Y1 - 1984
N2 - The nature of the proteoglycan(s) (PG) found in the extracellular matrix (ECM) layer produced by cultured bovine comeal endothelial (BCE) cells is analyzed. The PG(s) account for approximately 5 to 6% of the dry weight of the ECM, regardless of the amount of extracellular soluble PG available in the medium. A 4 M guanidinium chloride (GuCl) extract of ECM was separated on a dissociative cesium chloride (CsCl) gradient (1.25 g/cm3 starting density). Results showed one main peak of PG substance(s) comprising 91% of the total labelled substance and uronic acid, banding at a specific buoyant density of 1.29 g/cm3. The molecular weight of this major PG(s) as estimated by gel filtration on Sepharose CL-4B ranged from 0.5 to 0.7 × 106. Further chemical analysis of the main PG(s) band revealed a protein moiety accounting for 45% of the weight and carbohydrates-glycosaminoglycans (GAG) accounting for the remaining 55% Analysis of the GAG chains (over the entire gradient) showed a composition, based on the susceptibility of the PG substance(s) to degrading enzymes, of 50% heparan sulfate, 43.5% dermatan sulfate, and 6.5% chondroitin 4- and 6-sulfate chains. BCE cell cultures grown in the presence of βD-xyloside produced an ECM lacking more than 90% of the GAG content found in the control ECM. The medium-soluble GAG chains, produced in vast excess in cultures grown in the presence of βD-xyloside, are composed mainly of chondroitin 4- and 6-sulfates.
AB - The nature of the proteoglycan(s) (PG) found in the extracellular matrix (ECM) layer produced by cultured bovine comeal endothelial (BCE) cells is analyzed. The PG(s) account for approximately 5 to 6% of the dry weight of the ECM, regardless of the amount of extracellular soluble PG available in the medium. A 4 M guanidinium chloride (GuCl) extract of ECM was separated on a dissociative cesium chloride (CsCl) gradient (1.25 g/cm3 starting density). Results showed one main peak of PG substance(s) comprising 91% of the total labelled substance and uronic acid, banding at a specific buoyant density of 1.29 g/cm3. The molecular weight of this major PG(s) as estimated by gel filtration on Sepharose CL-4B ranged from 0.5 to 0.7 × 106. Further chemical analysis of the main PG(s) band revealed a protein moiety accounting for 45% of the weight and carbohydrates-glycosaminoglycans (GAG) accounting for the remaining 55% Analysis of the GAG chains (over the entire gradient) showed a composition, based on the susceptibility of the PG substance(s) to degrading enzymes, of 50% heparan sulfate, 43.5% dermatan sulfate, and 6.5% chondroitin 4- and 6-sulfate chains. BCE cell cultures grown in the presence of βD-xyloside produced an ECM lacking more than 90% of the GAG content found in the control ECM. The medium-soluble GAG chains, produced in vast excess in cultures grown in the presence of βD-xyloside, are composed mainly of chondroitin 4- and 6-sulfates.
UR - http://www.scopus.com/inward/record.url?scp=0021228689&partnerID=8YFLogxK
U2 - 10.3109/03008208409152142
DO - 10.3109/03008208409152142
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AN - SCOPUS:0021228689
SN - 0300-8207
VL - 13
SP - 45
EP - 57
JO - Connective Tissue Research
JF - Connective Tissue Research
IS - 1
ER -