Howard B V, Macarak E J, Gunson D, Kefalides N A
Proc Natl Acad Sci U S A. 1976 Jul;73(7):2361-4. doi: 10.1073/pnas.73.7.2361.
[14C]Proline and [14C]lysine were incorporated into collagen by cultures of endothelial cells derived from calf aortae. The isomer 3-hydroxy[14C]proline accounted for 10% of the total hydroxy[14C]proline in the collagen isolated from the medium. Approximately 81% of the hydroxy[14C]lysine isolated from the medium was glycosylated, and 91% of the glycosylated hydroxy[14C]lysine was in the form of the disaccharide glucosylgalactose. Gel filtration chromatography or acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate indicated that the initially synthesized peptide chain of [14C]collagen had a molecular weight of about 135,000; after pepsin digestion this was converted to 115,000. The ratio of hydroxy[14C]proline to total [14C]proline x 100 in the pesin-resistant fraction was 59. When examined by immunofluorescence microscopy, the endothelial cultures stained positively with antiserum to (Type IV) collagen from basement membrane of bovine anterior lens capsule. The data indicate that cultured endothelial cells derived from calf aortae synthesize collagen that resembles that of basement membrane collagen.
来自小牛主动脉的内皮细胞培养物将[14C]脯氨酸和[14C]赖氨酸掺入胶原蛋白中。从培养基中分离出的胶原蛋白中，异构体3-羟基[14C]脯氨酸占总羟基[14C]脯氨酸的10%。从培养基中分离出的羟基[14C]赖氨酸约81%被糖基化，且91%的糖基化羟基[14C]赖氨酸为二糖葡萄糖基半乳糖形式。在十二烷基硫酸钠存在下进行凝胶过滤色谱或丙烯酰胺凝胶电泳表明，最初合成的[14C]胶原蛋白肽链分子量约为135,000；经胃蛋白酶消化后，分子量转变为115,000。胃蛋白酶抗性组分中羟基[14C]脯氨酸与总[14C]脯氨酸的比例×100为59。通过免疫荧光显微镜检查时，内皮细胞培养物用针对牛晶状体前囊基底膜（IV型）胶原蛋白的抗血清染色呈阳性。数据表明，来自小牛主动脉的培养内皮细胞合成的胶原蛋白类似于基底膜胶原蛋白。
