Four new monomeric insulins obtained by alanine scanning the dimer-forming surface of the insulin molecule.
The residues A21Asn, B12Val, B16Tyr, B24Phe, B25Phe, B26Tyr and B27Thr, buried in the dimer of insulin, were identified by means of alanine-scanning mutagenesis. The receptor binding activity, in vivo biological potency and self-association properties of the seven single alanine human insulin mutants were determined. Four of the seven single alanine mutants, [B12Ala]human insulin, [B16Ala]human insulin, [B24Ala]human insulin and [B26Ala]human insulin, are monomeric insulin, which indicates that B12Val, B16Tyr, B24Phe and B26Tyr are crucial for the formation of insulin dimer. The monomeric [B16Ala]human insulin and [B26Ala]human insulin retain 27 and 54% receptor binding activity, respectively, and nearly the same in vivo biological potency compared with native insulin, so they could be developed as the fast-acting insulin.
Chen
H
H
State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Shi
M
M
Guo
Z Y
ZY
Tang
Y H
YH
Qiao
Z S
ZS
Liang
Z H
ZH
Feng
Y M
YM
D000409:Alanine / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular; D019281:Dimerization; D004926:Escherichia coli; D006801:Humans; D007328:Insulin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009154:Mutation; D011972:Receptor, Insulin / Q000378:metabolism
D007328:Insulin; D011972:Receptor, Insulin; D000409:Alanine