LABELED ADENOSINE(5')TETRAPHOSPHO(5')ADENOSINE (AP(4)A) AND ADENOSINE(5')TETRAPHOSPHO(5')NUCLEOSIDE (AP(4)N) - SYNTHESIS WITH FIREFLY LUCIFERASE

被引：7

作者：

GARRIDO, S ^{[1
]}

ZAERA, E ^{[1
]}

TORRECILLA, A ^{[1
]}

SILLERO, A ^{[1
]}

SILLERO, MAG ^{[1
]}

机构：

[1] UNIV AUTONOMA MADRID,FAC MED,CSIC,INST INVEST BIOMED,DEPT BIOQUIM,E-28029 MADRID,SPAIN

来源：

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 1995年 / 30卷 / 2-3期

关键词：

FIREFLY LUCIFERASE; AP(4)A; AP(4)N; DINUCLEOSIDE TETRAPHOSPHATE;

D O I：

10.1016/0165-022X(95)00007-E

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Labeled dinucleoside polyphosphates are not commercially available, in spite of being important molecules in metabolic regulation. Firefly luciferase (EC 1.13.12.7) is a useful enzyme for the synthesis of adenosine(5')tetraphospho(5')adenosine (Ap(4)A). As luciferase behaves as a nucleotidase at low ATP concentration, adequate concentrations (higher than 0.1 mM ATP) should be used to obtain a good yield of labeled Ap(4)A. [P-32]Ap(4)A has also been synthesized from ATP and [P-32]PPi. In a first step, [beta,gamma-P-32]ATP is generated in a ATP-[P-32]PPi exchange reaction catalyzed by luciferase. In a second step, the reaction is supplemented with pyrophosphatase and 32 P labeled Ap,A is obtained. Radioactive adenosine(5')tetraphospho(5')nucleoside (Ap(4)N) can also be synthesized from ATP gamma S and labeled NTP or from low concentrations of labeled ATP and high concentrations of cold NTP. The syntheses of radioactive Ap(n)A and p(n)A (n > 4) can also be approached with luciferase.

引用

页码：191 / 198

页数：8

共 15 条

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