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Modulating Insulin Fibrillation Using Engineered B-Chains with Mutated C-Termini.

Mohsen Akbarian1, Reza Yousefi1, Ali Akbar Moosavi-Movahedi2

  • 1Protein Chemistry Laboratory, Department of Biology, College of Sciences, Shiraz University, Shiraz, Iran.

Biophysical Journal
|October 15, 2019
PubMed
Summary
This summary is machine-generated.

Mutations in the insulin B-chain

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Chemistry

Background:

  • Insulin fibrillation is a significant challenge in medicine and biotechnology.
  • The precise molecular mechanisms of insulin fibrillation remain unclear.
  • Previous research implicated the insulin B-chain's C-terminal region in fibrillation.

Purpose of the Study:

  • To investigate the role of the insulin B-chain's C-terminal region in fibrillation.
  • To characterize the fibrillation kinetics and morphology of insulin mutants.
  • To explore potential applications of modified insulin analogs.

Main Methods:

  • Generation of T30R and K29R/T30R insulin B-chain mutants.
  • Recombinant production and combination of wild-type A-chain with mutant B-chains using chaperone αB-crystallin.
  • Comparative analysis of fibrillation kinetics, morphology, and driving forces for wild-type and mutant insulins.

Main Results:

  • Mutant insulins and B-chains showed enhanced resistance to stress-induced fibrillation, particularly during nucleation.
  • The B-chain appears to influence insulin fibrillation.
  • The A-chain plays a role in the lateral association of insulin fibrils, forming larger bundles.
  • The C-terminal region of the B-chain is crucial for fibrillation control and nucleus/protofibril formation.

Conclusions:

  • Both N-terminal and C-terminal regions of the insulin B-chain are critical regulators of fibrillation.
  • The C-terminal B-chain region likely acts as an attachment site for nucleus and protofibril formation.
  • The studied insulin variants exhibit improved stability and may serve as novel intermediate-acting insulin analogs for pharmaceutical use.