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Native Chemical Ligation to Minimize Aspartimide Formation during Chemical Synthesis of Small LDLa Protein
Author(s) -
Tailhades Julien,
Sethi Ashish,
Petrie Emma J.,
Gooley Paul R.,
Bathgate Ross A.,
Wade John D.,
Hossain Mohammed A.
Publication year - 2016
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201503599
Subject(s) - native chemical ligation , chemical ligation , chemistry , residue (chemistry) , combinatorial chemistry , peptide , chemical synthesis , cysteine , disulfide bond , chelation , recombinant dna , regioselectivity , stereochemistry , semisynthesis , peptide synthesis , biochemistry , organic chemistry , enzyme , in vitro , gene , catalysis
The inhibition of the G protein‐coupled receptor, relaxin family peptide receptor 1 (RXFP1), by a small LDLa protein may be a potential approach for prostate cancer treatment. However, it is a significant challenge to chemically produce the 41‐residue and three‐disulfide cross‐bridged LDLa module which is highly prone to aspartimide formation due to the presence of several aspartic acid residues. Known palliative measures, including addition of HOBt to piperidine for N α ‐deprotection, failed to completely overcome this side reaction. For this reason, an elegant native chemical ligation approach was employed in which two segments were assembled for generating the linear LDLa protein. Acquisition of correct folding was achieved by using either a regioselective disulfide bond formation or global oxidation strategies. The final synthetic LDLa protein obtained was characterized by NMR spectroscopic structural analysis after chelation with a Ca 2+ ion and confirmed to be equivalent to the same protein obtained by recombinant DNA production.