Semi-synthesis of a HGF/SF kringle one (K1) domain scaffold generates a potent in vivo MET receptor agonist

Abstract

Clustering of synthetic biotinylated K1 domain from HGF/SF by streptavidin is sufficient to generate a potent MET agonist.

Engineering growth factors for regenerative medicine applications

“…A yeast-displayed library of NK1 mutants was created by random mutagenesis and screened to identify variants with high levels of yeast cell surface expression compared to wild-type NK1; the best variants exhibited a 40-fold improved expression yield [31–33]. Improved soluble expression yields were also achieved by targeted mutagenesis of NK1 [30] or chemical synthesis of the K1 domains of NK1 [40], studies which are described in more detail below.…”

“…A biotin moiety was incorporated into this protein at the C-terminus and streptavidin was used to generate a multivalent and semisynthetic K1 construct (termed K1B/S). The K1B/S complex demonstrated potent agonistic activity in vitro , systemic activity in vivo , and efficient protection of Fas-induced liver apoptosis at doses 20-fold lower than full-length HGF [40]. …”

Accelerating chemoselective peptide bond formation using bis(2-selenylethyl)amido peptide selenoester surrogates

“…The assembly of the model peptide segments 10d , 19 and 22 involved the formation of two Gly–Cys junctions and yielded the 26 amino acids peptide 23d (27% overall after HPLC purification). The method was also applied to the synthesis of the 85 amino acids Kringle 1 domain (K1) of hepatocyte growth factor/scatter factor (HGF/SF) 42,71. This example involved the assembly of peptide segments 10c (HGF/SF 125–148), 18 (HGF/SF 149–176) and 21 (HGF/SF 177–209) through the sequential formation of Lys–Cys and Tyr–Cys junctions.…”

A simple and traceless solid phase method simplifies the assembly of large peptides and the access to challenging proteins

“…These are variants of the HGF Kringle 1 (K1) domain, which contains a high affinity binding site for HGF Receptor Tyrosine Kinase (RTK), also called MET. 51 The first segment equipped with the AcA group was immobilized on the hydroxylamine solid support as described before. The peptide chain was elongated in the N-to-C direction by activating the transformation of the SEA off group into a thioester with an excess of 3-mercaptopropionic acid (MPA) and then performing an NCL reaction in the presence of 4-mercaptophenylacetic acid (MPAA) as described previously.…”