Crystal structures of Candida albicans N-myristoyltransferase with two distinct inhibitors.
Myristoyl-CoA:protein N-myristoyltransferase (Nmt) is a monomeric enzyme that catalyzes the transfer of the fatty acid myristate from myristoyl-CoA to the N-terminal glycine residue of a variety of eukaryotic and viral proteins. Genetic and biochemical studies have established that Nmt is an attractive target for antifungal drugs. We present here crystal structures of C. albicans Nmt complexed with two classes of inhibitor competitive for peptide substrates. One is a peptidic inhibitor designed from the peptide substrate; the other is a nonpeptidic inhibitor having a benzofuran core. Both inhibitors are bound into the same binding groove, generated by some structural rearrangements of the enzyme, with the peptidic inhibitor showing a substrate-like binding mode and the nonpeptidic inhibitor binding differently. Further, site-directed mutagenesis for C. albicans Nmt has been utilized in order to define explicitly which amino acids are critical for inhibitor binding. The results suggest that the enzyme has some degree of flexibility for substrate binding and provide valuable information for inhibitor design.
Sogabe
Satoshi
S
Nippon Roche Research Center, 200 Kajiwara, Kamakura, Kanagawa 247-8530, Japan. satoshi.sogabe@roche.com
Masubuchi
Miyako
M
Sakata
Kiyoaki
K
Fukami
Takaaki A
TA
Morikami
Kenji
K
Shiratori
Yasuhiko
Y
Ebiike
Hirosato
H
Kawasaki
Kenichi
K
Aoki
Yuko
Y
Shimma
Nobuo
N
D'Arcy
Allan
A
Winkler
Fritz K
FK
Banner
David W
DW
Ohtsuka
Tatsuo
T
D000217:Acyltransferases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002176:Candida albicans / Q000201:enzymology*; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism; D007093:Imidazoles / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009842:Oligopeptides / Q000378:metabolism; D011487:Protein Conformation; D013379:Substrate Specificity
D000596:Amino Acids; D004791:Enzyme Inhibitors; D007093:Imidazoles; D009842:Oligopeptides; C104792:SC 58272; D000217:Acyltransferases; C050480:glycylpeptide N-tetradecanoyltransferase