Back to Search
Start Over
Native Chemical Ligation to Minimize Aspartimide Formation during Chemical Synthesis of Small LDLa Protein.
- Source :
-
Chemistry (Weinheim an der Bergstrasse, Germany) [Chemistry] 2016 Jan 18; Vol. 22 (3), pp. 1146-51. Date of Electronic Publication: 2015 Nov 27. - Publication Year :
- 2016
-
Abstract
- 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.<br /> (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Subjects :
- Adaptor Proteins, Vesicular Transport genetics
Amino Acid Sequence
Aspartic Acid chemistry
DNA, Recombinant genetics
Humans
Ligation
Magnetic Resonance Spectroscopy
Protein Binding
Receptors, G-Protein-Coupled chemistry
Receptors, G-Protein-Coupled genetics
Receptors, Peptide genetics
Adaptor Proteins, Vesicular Transport chemistry
Aspartic Acid analogs & derivatives
Calcium Chelating Agents chemistry
DNA, Recombinant chemistry
Receptors, G-Protein-Coupled antagonists & inhibitors
Receptors, Peptide chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1521-3765
- Volume :
- 22
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Chemistry (Weinheim an der Bergstrasse, Germany)
- Publication Type :
- Academic Journal
- Accession number :
- 26612092
- Full Text :
- https://doi.org/10.1002/chem.201503599