Isolation and characterisation of 5′‐fluorodeoxyadenosine synthase, a fluorination enzyme from Streptomyces cattleya

FEBS Letters

Deng, Hai¹  Schaffrath, Christoph¹  O'Hagan, David¹ 

[1] School of Chemistry and Centre for Biomolecular Sciences, University of St Andrews, North Haugh, St Andrews KY16 9ST, UK

关键词: 5′-Fluorodeoxyadenosine synthase;    S-Adenosyl-L-methionine;    Fluoride ion;    S-Adenosyl-L-homocysteine;    Streptomyces cattleya;    SAM;    S-adenosyl-L-methionine;    5′-FDA;    5′-fluoro-5′-deoxyadenosine;    SAH;    S-adenosyl-L-homocysteine;    ESI-MS;    electrospray ionisation mass spectrometry;   

DOI  :  10.1016/S0014-5793(03)00688-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

5′-Fluorodeoxyadenosine synthase, a C–F bond-forming enzyme, has been purified from Streptomyces cattleya. The enzyme mediates a reaction between inorganic fluoride and S-adenosyl-L-methionine (SAM) to generate 5′-fluoro-5′-deoxyadenosine. The molecular weight of the monomeric protein is shown to be 32.2 kDa by electrospray mass spectrometry. The kinetic parameters for SAM (K _m 0.42 mM, V _max 1.28 U/mg) and fluoride ion (K _m 8.56 mM, V _max 1.59 U/mg) have been evaluated. Both S-adenosyl-L-homocysteine (SAH) and sinefungin were explored as inhibitors of the enzyme. SAH emerged as a potent competitive inhibitor (K _i 29 μM) whereas sinefungin was only weakly inhibitory.

【 授权许可】

Unknown