Biomimetic synthesis of lispro insulin via a chemically synthesized "mini-proinsulin" prepared by oxime-forming ligation.
Here we report a proof-of-principle study demonstrating the efficient folding, with concomitant formation of the correct disulfides, of an isolated polypeptide insulin precursor of defined covalent structure. We used oxime-forming chemical ligation to introduce a temporary "chemical tether" to link the N-terminal residue of the insulin A chain to the C-terminal residue of the insulin B chain; the tether enabled us to fold/form disulfides with high efficiency. Enzymatic removal of the temporary chemical tether gave mature, fully active insulin. This chemical tethering principle could form the basis of a practical, high yield total synthesis of insulin and analogues.
Sohma
Youhei
Y
Department of Biochemistry and Molecular Biology, Institute for Biophysical Dynamics, The University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.
Kent
Stephen B H
SB
D004220:Disulfides; D007004:Hypoglycemic Agents / Q000138:chemical synthesis; D007328:Insulin / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D061268:Insulin Lispro; D010091:Oximes / Q000737:chemistry; D011384:Proinsulin / Q000138:chemical synthesis
D004220:Disulfides; D007004:Hypoglycemic Agents; D007328:Insulin; D061268:Insulin Lispro; D010091:Oximes; C060793:miniproinsulin; D011384:Proinsulin