Crystal structure of the anti-fungal target N-myristoyl transferase.
N-myristoyl transferase (NMT) catalyzes the transfer of the fatty acid myristate from myristoyl-CoA to the N-terminal glycine of substrate proteins, and is found only in eukaryotic cells. The enzyme in this study is the 451 amino acid protein produced by Candida albicans, a yeast responsible for the majority of systemic infections in immuno-compromised humans. NMT activity is essential for vegetative growth, and the structure was determined in order to assist in the discovery of a selective inhibitor of NMT which could be developed as an anti-fungal drug. NMT has no sequence homology with other protein sequences and has a novel alpha/beta fold which shows internal two-fold symmetry, which may be a result of gene duplication. On one face of the protein there is a long, curved, relatively uncharged groove, at the center of which is a deep pocket. The pocket floor is negatively charged due to the vicinity of the C-terminal carboxylate and a nearby conserved glutamic acid residue, which separates the pocket from a cavity. These observations, considered alongside the positions of residues whose mutation affects substrate binding and activity, suggest that the groove and pocket are the sites of substrate binding and the floor of the pocket is the catalytic center.
Weston
S A
SA
Zeneca Pharmaceuticals, Macclesfield, UK.
Camble
R
R
Colls
J
J
Rosenbrock
G
G
Taylor
I
I
Egerton
M
M
Tucker
A D
AD
Tunnicliffe
A
A
Mistry
A
A
Mancia
F
F
de la Fortelle
E
E
Irwin
J
J
Bricogne
G
G
Pauptit
R A
RA
D000214:Acyl Coenzyme A / Q000737:chemistry / Q000378:metabolism; D000217:Acyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D002177:Candidiasis; D018360:Crystallography, X-Ray; D005658:Fungi / Q000201:enzymology; D006801:Humans; D016867:Immunocompromised Host; D008958:Models, Molecular; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014978:Xenon
D000214:Acyl Coenzyme A; D000935:Antifungal Agents; D011994:Recombinant Proteins; C040349:S-tetradecanoyl-coenzyme A; D014978:Xenon; D000217:Acyltransferases; C050480:glycylpeptide N-tetradecanoyltransferase