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Modulating furin activity with designed mini‐PDX peptides: Synthesis and in vitro kinetic evaluation
Author(s) -
Basak Ajoy,
Lotfipour Farzaneh
Publication year - 2005
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.07.062
Subject(s) - furin , chemistry , peptide , proprotein convertases , biochemistry , serine , in vitro , tyrosine , cleavage (geology) , peptide sequence , mutant , enzyme , stereochemistry , biology , lipoprotein , paleontology , ldl receptor , cholesterol , fracture (geology) , gene
A peptide was designed from reactive site loop structure of α1 Antitrypsin Portland known as α1 PDX as a novel mini‐PDX inhibitor of furin. The sequence was derived from (367–394) that contains the crucial furin cleavage motif RIPR 382 . A P3 mutant replacing Ile 380 by Leu was prepared as a first model peptide. A Cys residue was inserted at each terminal of the peptide for purpose of cyclisation which was accomplished by air or iodine‐induced oxidation. This mini‐PDX peptide both cyclic and acyclic form inhibited in vitro furin activity (IC 50 in nM) when measured against either substrates Boc‐RVRR⇓MCA or QVEGF‐C [Abz‐QVHSIIRR⇓SLP‐Y(NO 2 )‐A‐CONH 2 , Abz = 2‐amino benzoic acid and Y(NO 2 ) = 3‐nitro tyrosine], latter being derived from vascular endothelial growth factor‐C (VEGF‐C) processing site. The geometrically constrained structure mimicking PDX reactive loop is crucial for enzyme inhibition. Our study further revealed that both mini‐PDX peptides inactivate furin in a slow tight binding manner, with disulfide‐bridged cyclic form being slightly more potent. Unlike PDX, these peptides inhibit furin via a different mechanistic pathway. The study provides an alternate strategy for development of efficient peptide‐based inhibitors of Proprotein Convertases including furin.