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

页码：164 / 171

页数：8

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[11] AN EFFICIENT METHOD FOR INVITRO LABELING PROTEINS TO HIGH SPECIFIC RADIOACTIVITY USING NABH-4-H-3 IN DIMETHYLFORMAMIDE [J].