Isolation and characterisation of 5'-fluorodeoxyadenosine synthase, a fluorination enzyme from Streptomyces cattleya.
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 microM) whereas sinefungin was only weakly inhibitory.
Schaffrath
Christoph
C
School of Chemistry and Centre for Biomolecular Sciences, University of St Andrews, North Haugh, KY16 9ST, St Andrews, UK.
Deng
Hai
H
O'Hagan
David
D
D000595:Amino Acid Sequence; D001711:Biotransformation; D007700:Kinetics; D008969:Molecular Sequence Data; D010430:Pentosyltransferases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry; D012436:S-Adenosylmethionine / Q000378:metabolism / Q000493:pharmacokinetics; D021241:Spectrometry, Mass, Electrospray Ionization; D013302:Streptomyces / Q000201:enzymology*
D010446:Peptide Fragments; D012436:S-Adenosylmethionine; C477004:5'-fluorodeoxyadenosine synthase; D010430:Pentosyltransferases