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Recombinant A17 Lys human insulin: purification and characterization
B Y Zhang1, Y S Zhang, R R Cai
1State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry, China.
Biochemistry and Molecular Biology International
|August 1, 1995
Summary
Recombinant A17 Lys human insulin showed lower receptor binding and biological activity compared to standard insulin. Researchers suggest the A17 glutamic acid residue is crucial for insulin
Area of Science:
- Biochemistry
- Molecular Biology
- Endocrinology
Background:
- Human insulin is a key hormone regulating blood glucose.
- Recombinant DNA technology allows for the production of modified insulin analogs.
- Understanding insulin structure-activity relationships is vital for diabetes treatment.
Purpose of the Study:
- To produce and characterize a novel human insulin analog with a lysine substitution at position A17.
- To evaluate the receptor-binding and biological activity of A17 Lys human insulin.
- To investigate the role of the A17 residue in insulin's function.
Main Methods:
- Expression of recombinant A17 Lys human insulin precursor in yeast.
- Isolation and purification of the insulin precursor from culture medium.
- Conversion of the precursor to A17 Lys human insulin.
- Crystallization and assessment of receptor-binding and biological activities.
Main Results:
- A17 Lys human insulin was successfully produced and purified.
- The A17 Lys analog could be crystallized.
- A17 Lys human insulin exhibited significantly lower receptor-binding affinity and biological activity compared to native human and porcine insulin.
- Native insulin contains glutamic acid at position A17, unlike the A17 Lys analog.
Conclusions:
- The A17 residue in human insulin plays a critical role in receptor binding and/or the active site.
- Substitution of A17 Glu with Lys significantly impairs insulin's biological function.
- Further studies are warranted to elucidate the precise mechanism of A17's involvement in insulin activity.