Turakainen H, Larjava H, Saarni H, Penttinen R
Biochim Biophys Acta. 1980 Apr 3;628(4):388-97. doi: 10.1016/0304-4165(80)90388-8.
Collagen and hyaluronic acid syntheses were studied in skin fibroblast cultures from patients with osteogenesis imperfecta and age-matched controls by labeling the cultures either with [3H]proline and separating the collagenous proteins with DEAE- and CM-cellulose chromatographies, or double-labeling the cultures with [3H]glucosamine and [14C]glycine and separating radioactive hyaluronic acid from glycoproteins and sulphated proteoglycans by DEAE-cellulose chromatography. The activities of the cell layer hyaluronate synthesizing enzyme complex (hyaluronate synthetase) were also determined. The osteogenesis imperfecta cultures were classified into three variants on the basis of type III collagen synthesis. Type III collagen amounted to approx. 40--50% from total collagen in the first variety and approx. 25--30% in the second variety. No difference was noted in the ratio of type III collagen to total collagen in the third variety in comparison with control cultures. The radioactivities of 3H-labeled hyaluronic acid in DEAE-cellulose chromatograms were compared with those of the 14C-labeled proteins. The ratios ranged 9.2--17.3 in the cultures from the patients and 4.6--8.8 in the control cultures. Hyaluronate synthetase activities were 1.3--2.0-fold higher in the osteogenesis imperfecta cells than in their controls. Increased hyaluronic acid synthesis in skin fibroblasts correlated with the severity of the disease but not with the increase in type III collagen synthesis.
通过用[3H]脯氨酸标记培养物并用DEAE - 纤维素和CM - 纤维素色谱法分离胶原蛋白质,或者用[3H]葡糖胺和[14C]甘氨酸对培养物进行双标记并用DEAE - 纤维素色谱法从糖蛋白和硫酸化蛋白聚糖中分离放射性透明质酸,研究了成骨不全患者和年龄匹配对照者皮肤成纤维细胞培养物中的胶原和透明质酸合成。还测定了细胞层透明质酸合成酶复合物(透明质酸合成酶)的活性。根据III型胶原合成情况,将成骨不全培养物分为三个变体。在第一个变体中,III型胶原约占总胶原的40 - 50%,在第二个变体中约占25 - 30%。与对照培养物相比,第三个变体中III型胶原与总胶原的比例没有差异。将DEAE - 纤维素色谱图中3H标记的透明质酸的放射性与14C标记的蛋白质的放射性进行比较。患者培养物中的比例范围为9.2 - 17.3,对照培养物中的比例范围为4.6 - 8.8。成骨不全细胞中的透明质酸合成酶活性比对照细胞高1.3 - 2.0倍。皮肤成纤维细胞中透明质酸合成增加与疾病严重程度相关,但与III型胶原合成增加无关。