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Structure-based stabilization of insulin as a therapeutic protein assembly via enhanced aromatic–aromatic interactions
- Source :
- Journal of Biological Chemistry. 293:10895-10910
- Publication Year :
- 2018
- Publisher :
- Elsevier BV, 2018.
-
Abstract
- Key contributions to protein structure and stability are provided by weakly polar interactions, which arise from asymmetric electronic distributions within amino acids and peptide bonds. Of particular interest are aromatic side chains whose directional π-systems commonly stabilize protein interiors and interfaces. Here, we consider aromatic–aromatic interactions within a model protein assembly: the dimer interface of insulin. Semi-classical simulations of aromatic–aromatic interactions at this interface suggested that substitution of residue Tyr(B26) by Trp would preserve native structure while enhancing dimerization (and hence hexamer stability). The crystal structure of a [Trp(B26)]insulin analog (determined as a T(3)R(f)(3) zinc hexamer at a resolution of 2.25 Å) was observed to be essentially identical to that of WT insulin. Remarkably and yet in general accordance with theoretical expectations, spectroscopic studies demonstrated a 150-fold increase in the in vitro lifetime of the variant hexamer, a critical pharmacokinetic parameter influencing design of long-acting formulations. Functional studies in diabetic rats indeed revealed prolonged action following subcutaneous injection. The potency of the Trp(B26)-modified analog was equal to or greater than an unmodified control. Thus, exploiting a general quantum-chemical feature of protein structure and stability, our results exemplify a mechanism-based approach to the optimization of a therapeutic protein assembly.
- Subjects :
- Male
Models, Molecular
0301 basic medicine
Protein Conformation
Dimer
Protein design
Random hexamer
Crystallography, X-Ray
Biochemistry
Diabetes Mellitus, Experimental
Amino Acids, Aromatic
03 medical and health sciences
Residue (chemistry)
chemistry.chemical_compound
Molecular dynamics
Protein structure
Animals
Insulin
Peptide bond
Amino Acid Sequence
Molecular Biology
chemistry.chemical_classification
030102 biochemistry & molecular biology
Cell Biology
Receptor, Insulin
Rats
Amino acid
030104 developmental biology
chemistry
Rats, Inbred Lew
Protein Structure and Folding
Biophysics
Dimerization
Protein Binding
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 293
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....1920c66b3e553caf1ac3c0983e89c908