AIM To study the relationship between insulinA chain regions and insulin biological activities,we designed a series of insulin analogues withchanges at A21,A12-18 of C-terminal helicalregion and A8-10 Iocated in the r...AIM To study the relationship between insulinA chain regions and insulin biological activities,we designed a series of insulin analogues withchanges at A21,A12-18 of C-terminal helicalregion and A8-10 Iocated in the region of A6-A11intra-chain disulphide bond.METHODS Insulin A-chain analogues wereprepared by stepwise Fmoc solid-phase manualsynthesis and then combined with natural B-chain of porcine insulin to yield correspondinginsulin analogues.Their biological activitieswere tested by receptor binding,mouseconvulsion and immunological assay.RESULTS[A21Ala]Ins retains 70.3% receptorbinding capacity and 60% in vivo biologicalactivity.[DesA13-14,A21Ala]Ins and[DesA12-13-14-15,A21Ala]Ins still have definite biologicalactivity,7.9% and 4.0% receptor binding,and6.2% and 3.3% in vivo biological activityrespectively.[A15Asn,A17Pro,A21Ala]Insmaintains 10.4% receptor binding and 10% invivo biological activity.[A8His,A9Arg,A10Pro,A21Ala]Ins,[A8His,A9Lys,A10Pro,A21Ala]Insand [A8His,A9Lys,A10Arg,A21Ala]Ins have51.9%,44.3% and 32.1% receptor bindingrespectively,50%,40% and 30% in vivobiological activity respectively,and 28.8%,29.6% and 15.4% immunological activityrespectively. CONCLUSION A21Asn can be replaced bysimple amino acid residues.The A chains withgradually damaged structural integrity in A12-18helical region and the demolition of the A12-18helical region by the substitution of Pro and Asnfor A17Glu and A15Gln respectively can combinewith the B chain and the combination productsshow definite biological activity,the helicalstructure of A12-18 is essential for biologicalactivities of insulin.A8-10 is not muchconcerned with biological activities,but is muchmore important antigenically in binding to itsantibodies,these results may help us design anew type of insulin analogue molecule.展开更多
基金the"Eighth Five Year Plan"Key Research Project,No.KS 852017National Natural Science Foundation of China.No.3880193,NO.39270157,No.39700028Chinese Academy of Sciences,KJ951-B1-606.
文摘AIM To study the relationship between insulinA chain regions and insulin biological activities,we designed a series of insulin analogues withchanges at A21,A12-18 of C-terminal helicalregion and A8-10 Iocated in the region of A6-A11intra-chain disulphide bond.METHODS Insulin A-chain analogues wereprepared by stepwise Fmoc solid-phase manualsynthesis and then combined with natural B-chain of porcine insulin to yield correspondinginsulin analogues.Their biological activitieswere tested by receptor binding,mouseconvulsion and immunological assay.RESULTS[A21Ala]Ins retains 70.3% receptorbinding capacity and 60% in vivo biologicalactivity.[DesA13-14,A21Ala]Ins and[DesA12-13-14-15,A21Ala]Ins still have definite biologicalactivity,7.9% and 4.0% receptor binding,and6.2% and 3.3% in vivo biological activityrespectively.[A15Asn,A17Pro,A21Ala]Insmaintains 10.4% receptor binding and 10% invivo biological activity.[A8His,A9Arg,A10Pro,A21Ala]Ins,[A8His,A9Lys,A10Pro,A21Ala]Insand [A8His,A9Lys,A10Arg,A21Ala]Ins have51.9%,44.3% and 32.1% receptor bindingrespectively,50%,40% and 30% in vivobiological activity respectively,and 28.8%,29.6% and 15.4% immunological activityrespectively. CONCLUSION A21Asn can be replaced bysimple amino acid residues.The A chains withgradually damaged structural integrity in A12-18helical region and the demolition of the A12-18helical region by the substitution of Pro and Asnfor A17Glu and A15Gln respectively can combinewith the B chain and the combination productsshow definite biological activity,the helicalstructure of A12-18 is essential for biologicalactivities of insulin.A8-10 is not muchconcerned with biological activities,but is muchmore important antigenically in binding to itsantibodies,these results may help us design anew type of insulin analogue molecule.
