ISOELECTRIC-FOCUSING OF SPARINGLY SOLUBLE-PROTEINS IN IMMOBILIZED PH GRADIENTS, EXEMPLIFIED BY MICROVILLAR MEMBRANE HYDROLASES

被引：29

作者：

SINHA, PK ^{[1
]}

RIGHETTI, PG ^{[1
]}

机构：

[1] FREE UNIV BERLIN, INST ANAT, D-1000 BERLIN 33, GERMANY

来源：

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 1986年 / 12卷 / 5-6期

关键词：

D O I：

10.1016/0165-022X(86)90065-5

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel fractionation technique is described for analysis of membrane-bound enzymes and sparingly soluble proteins: isoelectric focusing in a mixed-type matrix, containing a primary, immobilized pH gradient with a superimposed, secondary carrier ampholyte pH gradient. Three microvilli hydrolases; dipeptidyl peptidase IV, .gamma.-glutamyl transferase and alkaline phosphatase exhibit an array of sharply focused, enzyme active bands in the pH 4-6.5 range. The separation pattern obtained is by far superior to any separation achieved by either technique separately.

引用

页码：289 / 297

页数：9

共 26 条

[1] ISOELECTRIC-FOCUSING IN IMMOBILIZED PH GRADIENTS - PRINCIPLE, METHODOLOGY AND SOME APPLICATIONS [J].

BJELLQVIST, B ;

EK, K ;

RIGHETTI, PG ;

GIANAZZA, E ;

GORG, A ;

WESTERMEIER, R ;

POSTEL, W .

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 1982, 6 (04) :317-339

[2]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[3] PREPARATIVE ISOELECTRIC-FOCUSING IN IMMOBILIZED PH GRADIENTS .4. RECOVERY OF PROTEINS FROM IMMOBILINE MATRICES INTO ION-EXCHANGE RESINS [J].

CASERO, P ;

GELFI, C ;

RIGHETTI, PG .

ELECTROPHORESIS, 1985, 6 (02) :59-69

[4] ISOLATION AND CHARACTERIZATION OF THE PROXIMAL AND DISTAL FORMS OF LACTASE-PHLORIZIN HYDROLASE FROM THE SMALL-INTESTINE OF THE SUCKLING RAT [J].

COUSINEAU, J ;

GREEN, JR .

BIOCHIMICA ET BIOPHYSICA ACTA, 1980, 615 (01) :147-157

[5]

DANIELSEN EM, 1984, BIOCHEM J, V221, P1

[6] PERSPECTIVES ON ALKALINE-PHOSPHATASE ISOENZYMES [J].

FISHMAN, WH .

AMERICAN JOURNAL OF MEDICINE, 1974, 56 (05) :617-650

[7] SWELLING KINETICS OF IMMOBOILINE GELS FOR ISOELECTRIC-FOCUSING [J].

GELFI, C ;

RIGHETTI, PG .

ELECTROPHORESIS, 1984, 5 (05) :257-262

[8] PREPARATION OF IMMOBILIZED PH GRADIENTS SPANNING 2-6 PH UNITS WITH 2-CHAMBER MIXERS - EVALUATION OF 2 EXPERIMENTAL APPROACHES [J].

GIANAZZA, E ;

CELENTANO, F ;

DOSSI, G ;

BJELLQVIST, B ;

RIGHETTI, PG .

ELECTROPHORESIS, 1984, 5 (02) :88-97

[9] AMPHOLINE-AMPHOLINE INTERACTION AS A CAUSE OF PH GRADIENT DRIFT IN ISOELECTRIC-FOCUSING [J].

GIANAZZA, E ;

ASTORRI, C ;

RIGHETTI, PG .

JOURNAL OF CHROMATOGRAPHY, 1979, 171 (APR) :161-169

[10] PH, UREA AND SUBSTRATE GRADIENTS FOR THE OPTIMIZATION OF ULTRATHIN POLYACRYLAMIDE-GEL ZYMOGRAMS [J].

GORG, A ;

POSTEL, W ;

JOHANN, P .

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 1985, 10 (5-6) :341-350

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