HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY-BASED ASSAYS OF ENZYME-ACTIVITIES

被引:11
作者
LAMBETH, DO
MUHONEN, WW
机构
[1] Department of Biochemistry and Molecular Biology, School of Medicine, University of North Dakota, Grand Forks
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 1994年 / 656卷 / 01期
关键词
D O I
10.1016/0378-4347(94)00072-7
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Interest in using HPLC to assay enzymatic reactions continues to grow as evidenced by the more than 100 papers published during the early 1990s. HPLC can be used for any enzymatic assay that requires separation of substrates and products before quantifying the extent of the reaction. The popularity of HPLC-based assays is due to several reasons: (1) HPLC offers unsurpassed precision, specificity, sensitivity, and reproducibility. (2) Powerful microcomputers and user-friendly software automate the running of samples and collection and processing of data. (3) Current columns, especially C-18 packings, separate a very wide variety of samples, and (4) A variety of on-line detectors provide a means to detect virtually any compound. This review surveys recent papers on the development of HPLC-based assays for enzymes that degrade or otherwise modify macromolecules. Methods for assaying enzymes involved in metabolic pathways are also reviewed. Work by the authors in developing HPLC-based assays for mitochondrial enzymes that use GTP/GDP and other nucleotides that cannot be or are not easily assayed by enzyme-coupled assays is discussed. These enzymes include nucleoside diphosphate kinase, succinate thiokinase, and GTP-AMP phosphotransferase. The assays are suitable for determining the submitochondrial compartmentation of enzyme activities. Finally, current and anticipated trends in HPLC technology, including new column packings and the trend toward smaller columns that give faster separations, are reviewed in relation to enzyme assays.
引用
收藏
页码:143 / 157
页数:15
相关论文
共 113 条
[1]   HYDROPHILIC-INTERACTION CHROMATOGRAPHY FOR THE SEPARATION OF PEPTIDES, NUCLEIC-ACIDS AND OTHER POLAR COMPOUNDS [J].
ALPERT, AJ .
JOURNAL OF CHROMATOGRAPHY, 1990, 499 :177-196
[2]   SIMPLE ISOCRATIC HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR MEASURING PYRIDOXINE KINASE-ACTIVITY IN CRUDE BIOLOGICAL EXTRACTS [J].
ARGOUDELIS, CJ .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1990, 526 (01) :25-33
[3]   STUDIES OF THE POTENCY OF PROTEIN-KINASE INHIBITORS ON ATPASE ACTIVITIES [J].
BARRET, JM ;
ERNOULD, AP ;
ROUILLON, MH ;
FERRY, G ;
GENTON, A ;
BOUTIN, JA .
CHEMICO-BIOLOGICAL INTERACTIONS, 1993, 86 (01) :17-27
[4]   ASSAY OF PHENYLETHANOLAMINE N-METHYLTRANSFERASE ACTIVITY USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH ULTRAVIOLET ABSORBENCY DETECTION [J].
BEAUDOUIN, C ;
HAURAT, G ;
FRAISSE, L ;
SOUPPE, J ;
RENAUD, B .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1993, 613 (01) :51-58
[5]   APPLICATION OF HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY TO THE STUDY OF THIAMINE METABOLISM AND IN PARTICULAR THIAMINE TRIPHOSPHATASE [J].
BETTENDORFF, L .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1991, 566 (02) :397-408
[6]   HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF D-AMINO-ACID OXIDASE ACTIVITY [J].
BIONDI, PA ;
GUIDOTTI, L ;
NEGRI, A ;
SECCHI, C .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1991, 566 (02) :377-382
[7]   HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF THE METABOLITES OF NITRENDIPINE AND INVESTIGATION INTO THE METABOLIC PATHWAYS OF THIS DIHYDROPYRIDINE [J].
BOCKER, RH ;
PREUSS, E ;
PETER, R .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1990, 530 (01) :206-211
[8]   ASSAY FOR TRANS-PARA-COUMAROYL ESTERASE USING A SPECIFIC SUBSTRATE FROM PLANT-CELL WALLS [J].
BORNEMAN, WS ;
HARTLEY, RD ;
HIMMELSBACH, DS ;
LJUNGDAHL, LG .
ANALYTICAL BIOCHEMISTRY, 1990, 190 (01) :129-133
[9]   USE OF HYDROPHILIC INTERACTION CHROMATOGRAPHY FOR THE STUDY OF TYROSINE PROTEIN-KINASE SPECIFICITY [J].
BOUTIN, JA ;
ERNOULD, AP ;
FERRY, G ;
GENTON, A ;
ALPERT, AJ .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1992, 583 (02) :137-143
[10]   LONG-CHAIN ACYL-COA ESTER INTERMEDIATES OF BETA-OXIDATION OF MONO-CARBOXYLIC AND DI-CARBOXYLIC FATTY-ACIDS BY EXTRACTS OF CORYNEBACTERIUM SP STRAIN 7E1C [J].
BROADWAY, NM ;
DICKINSON, FM ;
RATLEDGE, C .
BIOCHEMICAL JOURNAL, 1992, 285 :117-122