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Decreased substrate affinity upon alteration of the substrate‐docking region in cytochrome P450 BM‐3
Author(s) -
Shelley A. Maves,
Hyeyeong Yeom,
Mark A. McLean,
Stephen G. Sligar
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00999-x
Subject(s) - chemistry , stereochemistry , mutant , kinetics , cytochrome p450 , substrate (aquarium) , docking (animal) , biochemistry , hydroxylation , amino acid , mutagenesis , saturated mutagenesis , glutamine , arginine , enzyme , proline , site directed mutagenesis , fatty acid , biology , gene , medicine , ecology , physics , nursing , quantum mechanics
A mutation at the surface of the substrate access channel which dramatically decreases the affinity for some fatty acids in P450 BM‐3 was discovered by random mutagenesis. The mutation introduced, proline‐25 to glutamine, is in close proximity to the arginine‐47 residue thought to be responsible for the initial docking of fatty acid substrates. The P25Q mutant displays an affinity for palmitate which is approximately 100‐fold weaker than the wild‐type enzyme. In addition to its altered substrate affinity, P25Q also exhibits altered hydroxylation specificity and carbon monoxide recombination kinetics in the substrate‐free form.