Studies on long-acting insulin: crystal structure of Arg-B31 human insulin at 2.0A resolution

The crystal structure of Arg-B31 human insulin (ABHI), a long-acting insulin derivative, has been determined at 2.0 A resolution by using X-ray diffraction analysis. The final crystallographic R factor of the structure model after the refinement is 0.189 with the bond length r. m. s deviation of 0.018 A. The refined structure of ABHI showed that the conformation of B-chain C-terminal residues was more stable than that in the native molecule. A striking structural feature of ABHI was an additional ion pair formed between Arg-B31 of molecule I and Glu-B21 of molecule II in a dimer, and three ionic bonds between the neighbouring molecules thereby appeared on the surface of ABHI hexamer. These secondary bonds generated by the insertion of the residue Arg-B31 should make the rate of dissociation of ABHI hexamer slow down when it was injected into the body and the property of protraction should be produced by a 'depot effect'. This ought to be the main structure basis of the prolonged action of ABHI. The results observed here demonstrate that the main idea we used in search for long-acting insulin is reasonable and correct which goes like this: making some additional non-covalent bonds between insulin monomers so as to slow down the dissociation of insulin oligomers and gain the protraction from a 'depot effect', which may be used as a principle in the further research. It also shows an impressive example that the experimental result reported here is in agreement with the theoretical prediction before the structural determination.