EC Number   |
|---|
   2.3.1.5 | - |
| 2.3.1.5 | crystal structure of MMNAT in complex with CoA is determined. The overall MMNAT structure consists of three domains. Domain I forms an alpha-helical bundle (amino acids 4-88), domain II forms a beta-barrel (amino acids 89-200) and domain III forms an alpha/beta lid (amino acids 201-275). Assembled together, the molecular surface of these domains presents a deep and wide active-site cleft, at the base of which is found the catalytic cysteine residue. Surprisingly, the principal CoA recognition site in MMNAT is located some 30 A from the site of CoA recognition in the deposited structure of human NAT2 bound to CoA |
| 2.3.1.5 | crystal structure of NfNAT is solved at a resolution of 2.7 A. Despite low sequence identity, enzyme shares an almost identical fold with that of other prokaryotic NATs, e.g. Mycobacterium smegmatis NAT and Mycobacterium marinum NAT. The overall NfNAT structure consists of three domains of equivalent size. The first two domains, a helical bundle (amino acids 1-98) and a beta-barrel (amino acids 99-199), are disposed in such a way that three residues (Cys82, His119, and Asp136) form a catalytic triad. The third domain is linked to the second through an interdomain helix (alpha6: amino acids 200-210). As shown for other prokaryotic NAT structures, the interface formed between domains 2 and 3 forms a substantial active-site cleft |
| 2.3.1.5 | crystallization in 0.5 M Ca(OAc)2, 16% PEG 3350, and 0.1 M Tris-HCl, pH 8.5, sitting drop vapor-diffusion method |
| 2.3.1.5 | crystallization of mutant Y183F is reported |
| 2.3.1.5 | crystals diffract to 1.95 A resolution on a synchrotron source |
| 2.3.1.5 | hanging drop vapor diffusion method, using 1.6 M sodium citrate, pH 6.5, 0.28 M NDSB-221 as additive |
| 2.3.1.5 | hanging- and sitting-drop vapour diffusion method |
| 2.3.1.5 | high resolution crystal structures of both human NATs, including NAT1 in complex with the irreversible inhibitor 2-bromoacetanilide, a site-directed NAT1 mutant, NAT1-F125S, and a NAT2-CoA complex are presented. By comparing the structures with known prokaryotic structures, evidences are provided for novel structural features of human NATs that are absent in bacterial enzymes, including an insertion that produces a significant difference in the structure of the carboxyl terminus of the eukaryotic enzymes |
| 2.3.1.5 | high resolution crystal structures of both human NATs, including NAT1 in complex with the irreversible inhibitor 2-bromoacetanilide, a site-directed NAT1 mutant, NAT1-F125S, and a NAT2-CoA complex are presented. By comparing the structures with known prokaryotic structures, evidences are provided for novel structural features of human NATs that are absent in bacterial enzymes, including an insertion that produces a significant difference in the structure of the carboxyl terminus of the eukaryotic enzymes. A complete picture of the distinct substrate selectivity of human NAT1 and NAT2, as well as key features of the naturally occurring variants of NAT1 and NAT2 is provided |
