Synthesis and biological activity of new iodoacetamide derivatives on mutants of squalene-hopene cyclase.
New iodoacetamide derivatives, containing a dodecyl or a squalenyl moiety, were synthesized. The effect of these new thiol-reacting molecules was studied on two mutants of Alicyclobacillus acidocaldarius squalene-hopene cyclase constructed especially for this purpose. In the quintuple mutant, all five cysteine residues of the enzyme are substituted with serine; in the sextuple mutant, this quintuple substitution is accompanied by the substitution of aspartate D376, located at the enzyme's active site, with a cysteine. N-Dodecyliodoacetamide had little activity toward either mutant, whereas N-squalenyliodoacetamide showed a stronger effect on the sextuple than on the quintuple mutant, as expected.
Ceruti
Maurizio
M
Dipartimento di Scienza e Tecnologia del Farmaco, Università degli Studi di Torino, 10125 Torino, Italy.
Balliano
Gianni
G
Rocco
Flavio
F
Lenhart
Alexander
A
Schulz
Georg E
GE
Castelli
Francesco
F
Milla
Paola
P
D001407:Bacillus / Q000201:enzymology; D019751:Intramolecular Transferases / Q000235:genetics / Q000378:metabolism*; D007460:Iodoacetamide / Q000138:chemical synthesis* / Q000494:pharmacology*; D009682:Magnetic Resonance Spectroscopy; D009154:Mutation; D021241:Spectrometry, Mass, Electrospray Ionization
D019751:Intramolecular Transferases; C066630:squalene-hopene cyclase; D007460:Iodoacetamide