A SPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF HEPARAN-SULFATE

被引：32

作者：

VANDELEST, CHA ^{[1
]}

VERSTEEG, EMM ^{[1
]}

VEERKAMP, JH ^{[1
]}

VANKUPPEVELT, TH ^{[1
]}

机构：

[1] CATHOLIC UNIV NIJMEGEN,DEPT BIOCHEM,6500 HB NIJMEGEN,NETHERLANDS

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 1994年 / 1201卷 / 02期

关键词：

HEPARAN SULFATE; GLYCOSAMINOGLYCAN; PROTEOGLYCAN; URINE; MUCOPOLYSACCHARIDOSIS;

D O I：

10.1016/0304-4165(94)90055-8

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：

A simple and reliable spectrophotometric method for the determination of heparan sulfate is described. The method is based on the 1,9-dimethylmethylene blue assay for sulfated glycosaminoglycans. Addition of bovine serum albumin, together with a specific NaCl concentration and pH, results in a specific decrease of heparan sulfate-based absorbance. The amount of heparan sulfate can be calculated by subtracting the values obtained in the presence of albumin from those obtained in its absence. The sensitivity is 0.5 mu g heparan sulfate. Two applications are given: the quantification of heparan sulfate in urine, including urine from patients with mucopolysaccharidosis, and the evaluation of fractions from gel filtration and ion exchange column chromatography for isolation of heparan sulfate proteoglycans.

引用

页码：305 / 311

页数：7

共 40 条

[1] CONFORMATIONAL FLEXIBILITY - A NEW CONCEPT FOR EXPLAINING BINDING AND BIOLOGICAL PROPERTIES OF IDURONIC ACID-CONTAINING GLYCOSAMINOGLYCANS [J].

CASU, B ;

PETITOU, M ;

PROVASOLI, M ;

SINAY, P .

TRENDS IN BIOCHEMICAL SCIENCES, 1988, 13 (06) :221-225

[2]

CELESIA GG, 1991, SEMIN THROMB HEMOST, V17, P158

[3] HUNTER-HURLER SYNDROME - GEL FILTRATION AND DIALYSIS OF URINARY ACID MUCOPOLYSACCHARIDES [J].

CONSTANTOPOULOS, G .

NATURE, 1968, 220 (5167) :583-+

[4]

DEJONG JGN, 1992, CLIN CHEM, V38, P803

[5] SELECTIVE URINARY SCREENING FOR MUCOPOLYSACCHARIDOSES [J].

DEMBURE, PP ;

DRUMHELLER, JE ;

BARR, SM ;

ELSAS, LJ .

CLINICAL BIOCHEMISTRY, 1990, 23 (01) :91-96

[6] IMPROVED QUANTITATION AND DISCRIMINATION OF SULFATED GLYCOSAMINOGLYCANS BY USE OF DIMETHYLMETHYLENE BLUE [J].

FARNDALE, RW ;

BUTTLE, DJ ;

BARRETT, AJ .

BIOCHIMICA ET BIOPHYSICA ACTA, 1986, 883 (02) :173-177

[7] A DIRECT SPECTROPHOTOMETRIC MICRO-ASSAY FOR SULFATED GLYCOSAMINOGLYCANS IN CARTILAGE CULTURES [J].

FARNDALE, RW ;

SAYERS, CA ;

BARRETT, AJ .

CONNECTIVE TISSUE RESEARCH, 1982, 9 (04) :247-248

[8] AN IMPROVED METHOD FOR DETERMINING PROTEOGLYCANS SYNTHESIZED BY CHONDROCYTES IN CULTURE [J].

GOLDBERG, RL ;

KOLIBAS, LM .

CONNECTIVE TISSUE RESEARCH, 1990, 24 (3-4) :265-275

[9] NOVEL AFFINITY CHROMATOGRAPHIC SYSTEM FOR THE SINGLE-STEP PURIFICATION OF GLYCOSAMINOGLYCANS FROM COMPLEX-SYSTEMS USING VOLATILE BUFFERS [J].

HODSON, BA ;

PEPPER, DS ;

DAWES, J .

JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1991, 565 (1-2) :416-423

[10]

HOFFMAN P, 1968, CHEM PHYSL MUCOPOLYS, P39

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