Aminodeoxychorismate synthase inhibitors from one-bead one-compound combinatorial libraries: "staged" inhibitor design.
4-Amino-4-deoxychorismate synthase (ADCS) catalyzes the first step in the conversion of chorismate into p-aminobenzoate, which is incorporated into folic acid. We aim to discover compounds that inhibit ADCS and serve as leads for a new class of antimicrobial compounds. This report presents (1) synthesis of a mass-tag encoded library based on a "staged" design, (2) massively parallel fluorescence-based on-bead screening, (3) rapid structural identification of hits, and (4) full kinetic analysis of ADCS. All inhibitors are competitive against chorismate and Mg(2+). The most potent ADCS inhibitor identified has a K(i) of 360 microM. We show that the combinatorial diversity elements add substantial binding affinity by interacting with residues outside of but proximal to the active site. The methods presented here constitute a paradigm for inhibitor discovery through active site targeting, enabled by rapid library synthesis, facile massively parallel screening, and straightforward hit identification.
Dixon
Seth
S
Department of Chemistry, University of California, One Shields Avenue, Davis, California 95616, USA.
Ziebart
Kristin T
KT
He
Ze
Z
Jeddeloh
Melissa
M
Yoo
Choong Leol
CL
Wang
Xiaobing
X
Lehman
Alan
A
Lam
Kit S
KS
Toney
Michael D
MD
Kurth
Mark J
MJ
D000085:Acetates / Q000138:chemical synthesis* / Q000737:chemistry; D000890:Anti-Infective Agents / Q000138:chemical synthesis* / Q000737:chemistry; D001565:Benzoates / Q000138:chemical synthesis* / Q000737:chemistry; D001665:Binding Sites; D019731:Carbon-Nitrogen Ligases / Q000037:antagonists & inhibitors* / Q000737:chemistry; D002413:Cations, Divalent; D002827:Chorismic Acid / Q000737:chemistry; D020650:Combinatorial Chemistry Techniques; D015195:Drug Design; D005456:Fluorescent Dyes; D062425:Hydroxybenzoate Ethers; D007700:Kinetics; D008274:Magnesium / Q000737:chemistry; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D010642:Phenoxyacetates / Q000138:chemical synthesis* / Q000737:chemistry; D011485:Protein Binding; D012117:Resins, Synthetic; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000637:Transaminases
C517235:3-(carboxymethoxy)benzoic acid; D000085:Acetates; D000890:Anti-Infective Agents; D001565:Benzoates; D002413:Cations, Divalent; D005456:Fluorescent Dyes; D062425:Hydroxybenzoate Ethers; D019151:Peptide Library; D010455:Peptides; D010642:Phenoxyacetates; D012117:Resins, Synthetic; D000637:Transaminases; C066599:aminodeoxychorismate synthase; D019731:Carbon-Nitrogen Ligases; D002827:Chorismic Acid; D008274:Magnesium