ENHANCEMENT OF FIREFLY LUCIFERASE ACTIVITY BY CYTIDINE NUCLEOTIDES

被引:29
作者
FORD, SR
HALL, MS
LEACH, FR
机构
[1] Department of Biochemistry, Oklahoma State University, Stillwater
关键词
D O I
10.1016/0003-2697(92)90239-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The temporal pattern of light production by firefly luciferase depends on the ATP concentration. With low concentrations of ATP a constant production of light occurred while at high concentrations of ATP (>10 μm) there was a flash of light followed by a decline in light production. This time course of light production with high ATP concentrations was changed from the flash pattern to a pattern with a constant production of light by several cytidine nucleotides. CTP, CDP, dCTP, dCDP, dideoxyCTP, periodate-oxidized CTP and CDP, and the etheno derivatives of CTP and CDP produced that change. CMP, cytidine, CDP-glycerol, CDP-glucose, CDP-ethanolamine, and benzoylbenzoylCTP either were inhibitory to firefly luciferase or were not effective in changing the flash time course. Coenzyme A and related compounds also changed the time course of light production. The changes in time course produced by either cytidine nucleotides or CoA were inhibited by desulfoCoA. These compounds apparently enhanced light production by promoting the dissociation of the inhibitory product, oxidized luciferin, from the enzyme. When the activating compounds were used with high concentrations of ATP, the sensitivity of assay for firefly luciferase was increased. This increased sensitivity is important when using the firefly luciferase gene as a reporter. © 1992.
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收藏
页码:283 / 291
页数:9
相关论文
共 51 条
[1]   THE FUNCTION OF COENZYME-A IN LUMINESCENCE [J].
AIRTH, RL ;
RHODES, WC ;
MCELROY, WD .
BIOCHIMICA ET BIOPHYSICA ACTA, 1958, 27 (03) :519-532
[2]   REPORTER GENES - APPLICATION TO THE STUDY OF MAMMALIAN GENE-TRANSCRIPTION [J].
ALAM, J ;
COOK, JL .
ANALYTICAL BIOCHEMISTRY, 1990, 188 (02) :245-254
[3]  
BRASIER AR, 1989, BIOTECHNIQUES, V7, P1116
[4]   AFFINITY LABELING OF PURINE NUCLEOTIDE SITES IN PROTEINS [J].
COLMAN, RF .
ANNUAL REVIEW OF BIOCHEMISTRY, 1983, 52 :67-91
[5]   FACTORS AFFECTING THE KINETICS OF LIGHT-EMISSION FROM CRUDE AND PURIFIED FIREFLY LUCIFERASE [J].
DELUCA, M ;
WANNLUND, J ;
MCELROY, WD .
ANALYTICAL BIOCHEMISTRY, 1979, 95 (01) :194-198
[6]   CONFORMATIONAL CHANGES OF LUCIFERASE DURING CATALYSES TRITIUM-HYDROGEN EXCHANGE AND OPTICAL ROTATION STUDIES [J].
DELUCA, M ;
MARSH, M .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1967, 121 (01) :233-&
[7]  
DELUCA M, 1984, BIOCHEM BIOPH RES CO, V123, P765
[8]  
DeLuca M., 1978, METHOD ENZYMOL, V57, P3, DOI DOI 10.1016/0076-6879(78)57003-1
[9]  
DeLuca M. A., 1981, BIOLUMINESCENCE CHEM
[10]  
DENBURG J L, 1970, Archives of Biochemistry and Biophysics, V141, P668, DOI 10.1016/0003-9861(70)90187-6