HIGH-PERFORMANCE AFFINITY-CHROMATOGRAPHY OF NADP+ DEHYDROGENASES FROM CELL-FREE-EXTRACTS USING A NUCLEOTIDE ANALOG AS GENERAL LIGAND

被引：4

作者：

ALHAMA, J ^{[1
]}

LOPEZBAREA, J ^{[1
]}

TORIBIO, F ^{[1
]}

机构：

[1] UNIV CORDOBA,FAC VET,DEPT BIOQUIM & BIOL MOLEC,AVDA MEDINA AZAHARA S-N,E-14071 CORDOBA,SPAIN

来源：

JOURNAL OF CHROMATOGRAPHY | 1991年 / 586卷 / 01期

关键词：

D O I：

10.1016/0021-9673(91)80024-B

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

An epoxy-activated silica column (50 cm x 0.45 cm I.D.) was derivatized with 8-[(6-aminohexyl)amino]-2'-phosphoadenosine-5'-diphosphoribose: the bound ligand concentration was 11.4-mu-mol g of dry silica, and the useful loading capacity was 2.3 mg of glutathione reductase. The new high-performance liquid chromatographic column specifically retained NADP+-dependent enzymes, which were quantitatively eluted specifically by NADP+ or with better resolution, by potassium chloride. The new high-performance liquid chromatographic support was applied to the purification of glutathione reductase and glucose-6-phosphate dehydrogenase from cell-free extracts of baker's yeast, fish liver and rabbit hemolysates, with high recoveries and excellent purification factors.

引用

页码：51 / 59

页数：9

共 36 条

[1] HIGH-PERFORMANCE LIQUID AFFINITY-CHROMATOGRAPHY WITH PHENYLBORONIC ACID, BENZAMIDINE, TRI-L-ALANINE, AND CONCANAVALIN-A IMMOBILIZED ON 3-ISOTHIOCYANATOPROPYLTRIETHOXYSILANE-ACTIVATED NONPOROUS MONODISPERSE SILICAS [J].

ANSPACH, FB ;

WIRTH, HJ ;

UNGER, KK ;

STANTON, P ;

DAVIES, JR ;

HEARN, MTW .

ANALYTICAL BIOCHEMISTRY, 1989, 179 (01) :171-181

[2]

ANSPACK B, 1988, J CHROMATOGR, V450, P195

[3] SIMPLE GENERAL METHOD FOR PREPARATION OF 6-IMMOBILIZED ANALOGS OF AMP, ATP, NAD AND OF OTHER ADENINE-CONTAINING COMPOUNDS FOR AFFINITY CHROMATOGRAPHY [J].

BARRY, S ;

OCARRA, P .

FEBS LETTERS, 1973, 37 (02) :134-139

[4]

BERGMEYER HU, 1974, METHOD ENZYMAT AN, V1, P443

[5]

BERGMEYER HU, 1974, METHOD ENZYMAT AN, V2, P575

[6]

BERGMEYER HU, 1974, METHOD ENZYMAT AN, V1, P510

[7] SYNTHESIS OF 3 AMP-ANALOGUES - N6-(6-AMINOHEXYL)-ADENOSINE 5'-MONOPHOSPHATE, N6-(6-AMINOHEXYL)-ADENOSINE 2',5'-BISPHOSPHATE, AND N6-(6-AMINOHEXYL)-ADENOSINE 3',5'-BISPHOSPHATE AND THEIR APPLICATION AS GENERAL LIGANDS IN BIOSPECIFIC AFFINITY CHROMATOGRAPH [J].

BRODELIUS, P ;

LARSSON, PO ;

MOSBACH, K .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1974, 47 (01) :81-89

[8] ANALYTICAL AFFINITY-CHROMATOGRAPHY IN STUDIES OF MOLECULAR RECOGNITION IN BIOLOGY - A REVIEW [J].

CHAIKEN, IM .

JOURNAL OF CHROMATOGRAPHY, 1986, 376 :11-32

[9]

CLONIS YD, 1986, J CHROMATOGR, V363, P31

[10] MATRIX EVALUATION FOR PREPARATIVE HIGH-PERFORMANCE AFFINITY-CHROMATOGRAPHY [J].

CLONIS, YD .

JOURNAL OF CHROMATOGRAPHY, 1987, 407 :179-187

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