ASSAY OF INTRACELLULAR THYMIDYLATE SYNTHETASE-ACTIVITY AND INHIBITION BY 5-FLUORO-2'-DEOXYURIDINE IN LYMPHOCYTES

被引：11

作者：

COX, S

HARMENBERG, J

机构：

[1] NATL BACTERIOL LAB,DEPT VIROL,S-10521 STOCKHOLM,SWEDEN

[2] KAROLINSKA INST,DEPT VIROL,S-10521 STOCKHOLM,SWEDEN

来源：

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 1992年 / 25卷 / 01期

关键词：

THYMIDYLATE SYNTHETASE; ASSAY; NUCLEOSIDE; NUCLEOSIDE ANALOG;

D O I：

10.1016/0165-022X(92)90032-6

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Thymidylate synthetase catalyses the formation of thymidine monophosphate from deoxyuridine monophosphate. Purified thymidylate synthetase can be assayed radiochemically using labelled deoxyuridine monophosphate as substrate, but cells are impervious to deoxyuridine monophosphate and so intracellular thymidylate synthetase activity cannot be assayed in this way. In this paper we describe the assay of intracellular thymidylate synthetase activity in intact cells using labelled 2'-deoxyuridine. The assay showed linear kinetics with respect to time, concentration of 2'-deoxyuridine, and cell concentration. 5-fluoro-2'-deoxyuridine inhibited intracellular thymidylate synthetase activity measured with this assay by 50% at 5 nM. Cell growth was inhibited by 50% at 6 nM 5-fluoro-2'-deoxyuridine. The assay was specific for thymidylate synthetase and enabled measurement of thymidylate synthetase activity in situ in intact cells.

引用

页码：17 / 23

页数：7

共 50 条

[21] INTERACTION OF PURIFIED HUMAN THYMIDYLATE SYNTHETASE (TSASE) WITH 5-FLUORO-2'-DEOXYURIDYLATE (FDUMP)
LOCKSHIN, A
DANENBERG, PV
PROCEEDINGS OF THE AMERICAN ASSOCIATION FOR CANCER RESEARCH, 1979, 20 (MAR): : 83 - 83
[22] 3',5'-DIESTERS OF 5-FLUORO-2'-DEOXYURIDINE - SYNTHESIS AND BIOLOGICAL ACTIVITY
NISHIZAWA, Y
CASIDA, JE
ANDERSON, SW
HEIDELBE.C
BIOCHEMICAL PHARMACOLOGY, 1965, 14 (11) : 1605 - +
[23] SYNTHESIS AND BIOLOGICAL-ACTIVITY OF 5-FLUORO-2'-DEOXYURIDINE 5'-PHOSPHORODIAMIDATES
PHELPS, ME
WOODMAN, PW
DANENBERG, PV
JOURNAL OF MEDICINAL CHEMISTRY, 1980, 23 (11) : 1229 - 1232
[24] Synthesis and anticancer activity of some 5-fluoro-2′-deoxyuridine phosphoramidates
Lewandowska, Marta
Ruszkowski, Piotr
Chojnacka, Kinga
Kleczewska, Natalia
Hoffmann, Marcin
Kacprzak, Karol
Celewicz, Lech
BIOORGANIC & MEDICINAL CHEMISTRY, 2016, 24 (10) : 2330 - 2341
[25] Discovery of 5′-Substituted 5-Fluoro-2′-deoxyuridine Monophosphate Analogs: A Novel Class of Thymidylate Synthase Inhibitors
Dasari, Madhuri
Pelly, Stephen C.
Geng, Jiafeng
Gold, Hannah B.
Pribut, Nicole
Sharma, Savita K.
D'Erasmo, Michael P.
Bartsch, Perry W.
Sun, Carrie
Toti, Kiran
Arnold, Rebecca S.
Petros, John A.
Xu, Lingjie
Jiang, Yi
Liotta, Dennis C.
Miller, Eric J.
ACS PHARMACOLOGY & TRANSLATIONAL SCIENCE, 2023, 6 (05) : 702 - 709
[26] THERMODYNAMIC ANALYSIS OF THE BINDING OF 5-FLUORO-2'-DEOXYURIDINE 5'-MONOPHOSPHATE TO THYMIDYLATE SYNTHASE OVER A RANGE OF TEMPERATURES
GARCIAFUENTES, L
RECHE, P
LOPEZMAYORGA, O
SANTI, DV
GONZALEZPACANOWSKA, D
BARON, C
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1995, 232 (02): : 641 - 645
[27] THYMIDYLATE SYNTHASE FROM UNTREATED HUMAN COLORECTAL-CANCER AND COLONIC MUCOSA - ENZYME-ACTIVITY AND INHIBITION BY 5-FLUORO-2'-DEOXYURIDINE-5'-MONOPHOSPHATE
PETERS, GJ
VANGROENINGEN, CJ
LAURENSSE, EJ
PINEDO, HM
EUROPEAN JOURNAL OF CANCER, 1991, 27 (03) : 263 - 267
[28] CYTOTOXICITY OF 5-NITRO-2'-DEOXYURIDINE BY INVIVO INHIBITION OF THYMIDYLATE SYNTHETASE
WASHTIEN, W
MATSUDA, A
WATAYA, Y
SANTI, DV
BIOCHEMICAL PHARMACOLOGY, 1978, 27 (22) : 2663 - 2666
[29] INHIBITION OF MAMMALIAN TUMOR THYMIDYLATE SYNTHETASE BY 2'-DEOXYURIDINE 5'-PHOSPHATE ANALOGS
RODE, W
KULIKOWSKI, T
KEDZIERSKA, B
SHUGAR, D
DRUGS UNDER EXPERIMENTAL AND CLINICAL RESEARCH, 1986, 12 (6-7) : 545 - 549
[30] ANTITUMOR-ACTIVITY OF ALKYLESTERS OF 5-FLUORO-2'-DEOXYURIDINE 5'-MONOPHOSPHATE (FDUMP) AGAINST MURINE LYMPHOMA L5178Y RESISTANT TO 5-FLUORO-2'-DEOXYURIDINE
KANZAWA, F
MATSUSHIMA, Y
ISHIHARA, JI
HOSHI, A
OHBA, T
WATANABE, K
JOURNAL OF PHARMACOBIO-DYNAMICS, 1986, 9 (08): : 688 - 693

← 1 2 3 4 5 →