2JZQ

Design of an Active Ultra-Stable Single-Chain Insulin Analog 20 Structures

Summary for 2JZQ

• Entry DOI: 10.2210/pdb2jzq/pdb
• Descriptor: Insulin (1 entity in total)
• Functional Keywords: hormone, single chain insulin, mutant, carbohydrate metabolism, cleavage on pair of basic residues, diabetes mellitus, disease mutation, glucose metabolism, pharmaceutical, secreted
• Cellular location: Secreted: P01308
• Total number of polymer chains: 1
• Total formula weight: 6382.19

Authors

Hua, Q.X.,Nakarawa, S.,Jia, W.H.,Huang, K.,Philips, N.F.,Hu, S.Q.,Weiss, M.A. (deposition date: 2008-01-11, release date: 2008-02-26, Last modification date: 2024-10-30)

Primary citation

Hua, Q.X.,Nakagawa, S.H.,Jia, W.,Huang, K.,Phillips, N.B.,Hu, S.Q.,Weiss, M.A.
Design of an active ultrastable single-chain insulin analog: synthesis, structure, and therapeutic implications.
J.Biol.Chem., 283:14703-14716, 2008

PubMed Abstract: Single-chain insulin (SCI) analogs provide insight into the inter-relation of hormone structure, function, and dynamics. Although compatible with wild-type structure, short connecting segments (<3 residues) prevent induced fit upon receptor binding and so are essentially without biological activity. Substantial but incomplete activity can be regained with increasing linker length. Here, we describe the design, structure, and function of a single-chain insulin analog (SCI-57) containing a 6-residue linker (GGGPRR). Native receptor-binding affinity (130 +/- 8% relative to the wild type) is achieved as hindrance by the linker is offset by favorable substitutions in the insulin moiety. The thermodynamic stability of SCI-57 is markedly increased (DeltaDeltaG(u) = 0.7 +/- 0.1 kcal/mol relative to the corresponding two-chain analog and 1.9 +/- 0.1 kcal/mol relative to wild-type insulin). Analysis of inter-residue nuclear Overhauser effects demonstrates that a native-like fold is maintained in solution. Surprisingly, the glycine-rich connecting segment folds against the insulin moiety: its central Pro contacts Val(A3) at the edge of the hydrophobic core, whereas the final Arg extends the A1-A8 alpha-helix. Comparison between SCI-57 and its parent two-chain analog reveals striking enhancement of multiple native-like nuclear Overhauser effects within the tethered protein. These contacts are consistent with wild-type crystal structures but are ordinarily attenuated in NMR spectra of two-chain analogs, presumably due to conformational fluctuations. Linker-specific damping of fluctuations provides evidence for the intrinsic flexibility of an insulin monomer. In addition to their biophysical interest, ultrastable SCIs may enhance the safety and efficacy of insulin replacement therapy in the developing world.

PubMed: 18332129
DOI: 10.1074/jbc.M800313200

Experimental method

SOLUTION NMR