A HIGHLY SPECIFIC AND QUANTITATIVE METHOD FOR DETERMINING TYPE-III-I COLLAGEN RATIOS IN TISSUES

被引:27
作者
CHEUNG, DT
BENYA, PD
PERELMAN, N
DICESARE, PE
NIMNI, ME
机构
[1] UNIV SO CALIF,SCH MED,DEPT BIOCHEM,LOS ANGELES,CA 90007
[2] UNIV SO CALIF,SCH MED,DEPT MED,LOS ANGELES,CA 90007
来源
MATRIX | 1990年 / 10卷 / 03期
关键词
CNBr peptides; collagen; skin; type I; type III;
D O I
10.1016/S0934-8832(11)80165-4
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The distribution of type I and type III collagens in rat, bovine and human skin were examined by a quantitative 2-D CNBr peptide mapping method.The procedure involved the solubilization of tissues by digestion with CNBr, radioactive labeling in vitro by [3H]-NaBH4 in dimethylformamide, reduction by mercaptoethanol, a second CNBr digestion and 2-D (isoelectric focusing and NaDodS04 electrophoresis) mapping.The amounts of type I and type III collagen peptide spots in the fluorographs of 2-D maps were analyzed by 2-D scanning densitometer/analyzer.Mixtures containing various ratios of purified type I and type III collagen were used to obtain a standard curve.Using this procedure we were able to determine that in adult human skin (age range 35 - 65 years) 22 % (+ 1.3 %) of the labelled collagen is type III.This value is significantly higher than that was previously estimated by less accurate methods. © 1990, Gustav Fischer Verlag · Stuttgart · New York. All rights reserved.
引用
收藏
页码:164 / 171
页数:8
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