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Design and synthesis of substrate‐based inhibitors of botulinum neurotoxin type B metalloprotease
Author(s) -
Oost Thorsten,
Sukonpan Chanokporn,
Brewer Matthias,
Goodnough Michael,
Tepp William,
Johnson Eric A.,
Rich Daniel H.
Publication year - 2003
Publication title -
peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.10590
Subject(s) - chemistry , dipeptide , peptide , stereochemistry , proteases , amide , cleavage (geology) , substrate (aquarium) , tetrahedral carbonyl addition compound , transition state analog , peptide bond , enzyme , active site , biochemistry , nucleophile , oceanography , geotechnical engineering , fracture (geology) , engineering , geology , catalysis
Botulinum toxin (BoNT) metalloproteases and related proteases are the most selective proteases known. X‐ray crystal structures suggest that the native enzymes exist in catalytically incompetent forms that must be activated by substrate binding. In order to characterize the postulated substrate‐induced conformational changes, we synthesized a series of transition state analog inhibitors (TSI) in which the dipeptide cleavage site has been replaced by tetrahedral intermediate analogs within the minimal substrate peptide sequence. Reduced amide, α‐hydroxyamide, α‐thio‐amide, and hydroxyethylamine analogs of —Gln–Phe— were incorporated via solid phase peptide synthesis into 35‐mer analogs of the minimal peptide substrate sequence. The synthesis, characterization, and inhibition kinetics for four series of compounds against holotoxin BoNT/B is described. The α‐thiol amide derivatives of the 35‐mer substrate were found to inhibit BONT/B in the low micromolar range. © 2003 Wiley Periodicals, Inc. Biopolymers (Pept Sci), 2003