SPECIFIC PROTEIN-BINDING REACTIONS MONITORED WITH LIGAND-ATP CONJUGATES AND FIREFLY LUCIFERASE

被引：25

作者：

CARRICO, RJ ^{[1
]}

YEUNG, KK ^{[1
]}

SCHROEDER, HR ^{[1
]}

BOGUSLASKI, RC ^{[1
]}

BUCKLER, RT ^{[1
]}

CHRISTNER, JE ^{[1
]}

机构：

[1] AMES CO, MILES LABS INC, ELKHART, IN 46514 USA

来源：

ANALYTICAL BIOCHEMISTRY | 1976年 / 76卷 / 01期

关键词：

D O I：

10.1016/0003-2697(76)90267-0

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A new method was developed to monitor specific protein binding reactions with an ATP-labeled ligand and firefly luciferase. The ligand 2,4-dinitrobenzene was covalently coupled to 4 ATP derivatives, and 3 of these conjugates were measured quantitatively at nanomolar levels with firefly luciferase. Incubation of the conjugates with antibody to the 2,4-dinitrophenyl residue diminished the peak light intensities produced in the bioluminescent assay; incubation with immunoglobulin from a nonimmunized rabbit did not affect light production. The antibody-bound ligand-ATP conjugates were inactive in the bioluminescent assay, and levels of unbound conjugate could be measured in the presence of the bound form. The firefly luciferase was used to monitor competitive binding reactions between the antibody, the conjugates and N(2,4-dinitrophenyl)-.beta.-alanine.

引用

页码：95 / 110

页数：16

共 18 条

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