Supramolecular Protein Engineering: DESIGN OF ZINC-STAPLED INSULIN HEXAMERS AS A LONG ACTING DEPOT

Bottom-up control of supramolecular protein assembly can provide a therapeutic nanobiotechnology. We demonstrate that the pharmacological properties of insulin can be enhanced by design of "zinc staples" between hexamers. Paired (i, i+4) His substitutions were introduced at an α-helical su...

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Bibliographic Details
Published in:The Journal of biological chemistry 2010-04, Vol.285 (16), p.11755-11759
Main Authors: Phillips, Nelson B, Wan, Zhu-li, Whittaker, Linda, Hu, Shi-Quan, Huang, Kun, Hua, Qing-xin, Whittaker, Jonathan, Ismail-Beigi, Faramarz, Weiss, Michael A
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Binding Sites
Crystallography, X-Ray
Delayed-Action Preparations
Drug Design
Humans
Insulin - administration & dosage
Insulin - analogs & derivatives
Insulin - chemical synthesis
Insulin - chemistry
Models, Molecular
Molecular Sequence Data
Nuclear Magnetic Resonance, Biomolecular
Protein Engineering - methods
Protein Structure and Folding
Protein Structure, Secondary
Rats
Static Electricity
Zinc - chemistry
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Summary:Bottom-up control of supramolecular protein assembly can provide a therapeutic nanobiotechnology. We demonstrate that the pharmacological properties of insulin can be enhanced by design of "zinc staples" between hexamers. Paired (i, i+4) His substitutions were introduced at an α-helical surface. The crystal structure contains both classical axial zinc ions and novel zinc ions at hexamer-hexamer interfaces. Although soluble at pH 4, the combined electrostatic effects of the substitutions and bridging zinc ions cause isoelectric precipitation at neutral pH. Following subcutaneous injection in a diabetic rat, the analog effected glycemic control with a time course similar to that of long acting formulation Lantus®. Relative to Lantus, however, the analog discriminates at least 30-fold more stringently between the insulin receptor and mitogenic insulin-like growth factor receptor. Because aberrant mitogenic signaling may be associated with elevated cancer risk, such enhanced specificity may improve safety. Zinc stapling provides a general strategy to modify the pharmacokinetic and biological properties of a subcutaneous protein depot.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.C110.105825