Backbone cyclic insulin

Backbone cyclic insulin was designed and prepared by reverse proteolysis in partial organic solvent of a single‐chain precursor expressed in yeast. The precursor contains two loops to bridge the two chains of native insulin. The cyclisation method uses Achromobacter lyticus protease and should be ge...

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Bibliographic Details
Published in:Journal of peptide science 2010-09, Vol.16 (9), p.473-479
Main Authors: Andersen, Asser S., Palmqvist, Eva, Bang, Susanne, Shaw, Allan C., Hubalek, Frantisek, Ribel, Ulla, Hoeg-Jensen, Thomas
Format: Article
Language:English
Subjects:
Acetylcholine receptors
Achromobacter
Achromobacter lyticus protease
Amino Acid Sequence
Animals
carboxypeptidase Y
Chromatography, Liquid
Chymotrypsin
cyclic
Cyclization
Insulin
Insulin - analogs & derivatives
Insulin - chemistry
Insulin - pharmacology
Insulin receptors
Intravenous administration
Lysine
Mass Spectrometry
Molecular Sequence Data
Peptides, Cyclic - chemical synthesis
Peptides, Cyclic - pharmacology
Proteinase
Proteolysis
Rats
Rats, Wistar
Receptor, Insulin - metabolism
semi-synthesis
Solvents
Wistar rats
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Summary:Backbone cyclic insulin was designed and prepared by reverse proteolysis in partial organic solvent of a single‐chain precursor expressed in yeast. The precursor contains two loops to bridge the two chains of native insulin. The cyclisation method uses Achromobacter lyticus protease and should be generally applicable to proteins with C‐terminal lysine and proximal N‐terminal. The presence of the ring‐closing bond and the native insulin disulfide patterns were documented by LC–MS peptide maps. The cyclic insulin was shown to be inert towards degradation by CPY, but was somewhat labile towards chymotrypsin. Intravenous administration of the cyclic insulin to Wistar rats showed the compounds to be equipotent to HI despite much lower insulin receptor affinity. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd. Backbone cyclic insulin was engineered from a yeast‐based expression system via addition of two loops and the cyclisation bond formed by reverse proteolysis, providing carboxypeptidase inert cyclic insulin.
ISSN:1075-2617
1099-1387
1099-1387
DOI:10.1002/psc.1264