PDBsum entry 1hit

Go to PDB code: 
protein Protein-protein interface(s) links
Hormone PDB id
Protein chains
21 a.a.
30 a.a. *
* Residue conservation analysis
PDB id:
Name: Hormone
Title: Receptor binding redefined by a structural switch in a mutan insulin
Structure: Insulin. Chain: a. Engineered: yes. Insulin. Chain: b. Engineered: yes. Mutation: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Organism_taxid: 9606
NMR struc: 9 models
Authors: Q.X.Hua,M.Kochoyan,M.A.Weiss
Key ref: Q.X.Hua et al. (1991). Receptor binding redefined by a structural switch in a mutant human insulin. Nature, 354, 238-241. PubMed id: 1961250
28-Feb-92     Release date:   31-Jan-94    
Go to PROCHECK summary

Protein chain
Pfam   ArchSchema ?
P01308  (INS_HUMAN) -  Insulin
110 a.a.
21 a.a.
Protein chain
Pfam   ArchSchema ?
P01308  (INS_HUMAN) -  Insulin
110 a.a.
30 a.a.*
Key:    PfamA domain  Secondary structure
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     extracellular region   1 term 
  Biochemical function     hormone activity     1 term  


Nature 354:238-241 (1991)
PubMed id: 1961250  
Receptor binding redefined by a structural switch in a mutant human insulin.
Q.X.Hua, S.E.Shoelson, M.Kochoyan, M.A.Weiss.
Crystal structures of insulin have been determined in various distinct forms, the relevance of which to receptor recognition has long been the subject of speculation. Recently the crystal structure of an inactive insulin analogue has been determined and, surprisingly, found to have a conformation identical to native insulin. On this basis Dodson and colleagues have suggested that the known insulin crystal structures reflect an inactive conformation, and that a change in conformation is required for activity--specifically, the carboxy terminal residues of the B-chain are proposed to separate from the amino terminal residues of the A-chain. Here we report the solution structure of an active insulin mutant, determined by two-dimensional NMR, which supports this hypothesis. In the mutant, the carboxy terminal beta-turn and beta-strand of the B-chain are destabilized and do not pack across the rest of the molecule. We suggest that analogous detachment of the carboxy terminal region of the B-chain occurs in native insulin on binding to its receptor. Our finding that partial unfolding of the B-chain exposes an alternative protein surface rationalizes the receptor-binding properties of a series of anomalous insulin analogues, including a mutant insulin associated with diabetes mellitus in man.

Literature references that cite this PDB file's key reference

  PubMed id Reference
23302862 J.G.Menting, J.Whittaker, M.B.Margetts, L.J.Whittaker, G.K.Kong, B.J.Smith, C.J.Watson, L.Záková, E.Kletvíková, J.Jiráček, S.J.Chan, D.F.Steiner, G.G.Dodson, A.M.Brzozowski, M.A.Weiss, C.W.Ward, and M.C.Lawrence (2013).
How insulin engages its primary binding site on the insulin receptor.
  Nature, 493, 241-245.
PDB codes: 3w11 3w12 3w13 3w14
20133841 J.Jirácek, L.Záková, E.Antolíková, C.J.Watson, J.P.Turkenburg, G.G.Dodson, and A.M.Brzozowski (2010).
Implications for the active form of human insulin based on the structural convergence of highly active hormone analogues.
  Proc Natl Acad Sci U S A, 107, 1966-1970.
PDB codes: 2wru 2wrv 2wrw 2wrx 2ws0 2ws1 2ws4 2ws6 2ws7
20208353 K.Nagata (2010).
Studies of the structure-activity relationships of peptides and proteins involved in growth and development based on their three-dimensional structures.
  Biosci Biotechnol Biochem, 74, 462-470.  
19321435 B.Xu, K.Huang, Y.C.Chu, S.Q.Hu, S.Nakagawa, S.Wang, R.Y.Wang, J.Whittaker, P.G.Katsoyannis, and M.A.Weiss (2009).
Decoding the Cryptic Active Conformation of a Protein by Synthetic Photoscanning: INSULIN INSERTS A DETACHABLE ARM BETWEEN RECEPTOR DOMAINS.
  J Biol Chem, 284, 14597-14608.  
19139090 C.L.Alvino, K.A.McNeil, S.C.Ong, C.Delaine, G.W.Booker, J.C.Wallace, J.Whittaker, and B.E.Forbes (2009).
A Novel Approach to Identify Two Distinct Receptor Binding Surfaces of Insulin-like Growth Factor II.
  J Biol Chem, 284, 7656-7664.  
19274663 C.W.Ward, and M.C.Lawrence (2009).
Ligand-induced activation of the insulin receptor: a multi-step process involving structural changes in both the ligand and the receptor.
  Bioessays, 31, 422-434.  
19883595 I.B.Bekard, and D.E.Dunstan (2009).
Tyrosine autofluorescence as a measure of bovine insulin fibrillation.
  Biophys J, 97, 2521-2531.  
19533727 J.Haas, E.Vöhringer-Martinez, A.Bögehold, D.Matthes, U.Hensen, A.Pelah, B.Abel, and H.Grubmüller (2009).
Primary steps of pH-dependent insulin aggregation kinetics are governed by conformational flexibility.
  Chembiochem, 10, 1816-1822.  
19321436 Q.X.Hua, B.Xu, K.Huang, S.Q.Hu, S.Nakagawa, W.Jia, S.Wang, J.Whittaker, P.G.Katsoyannis, and M.A.Weiss (2009).
Enhancing the Activity of a Protein by Stereospecific Unfolding: CONFORMATIONAL LIFE CYCLE OF INSULIN AND ITS EVOLUTIONARY ORIGINS.
  J Biol Chem, 284, 14586-14596.
PDB codes: 2k91 2k9r
19086273 E.E.Büllesbach, M.A.Hass, M.R.Jensen, D.F.Hansen, S.M.Kristensen, C.Schwabe, and J.J.Led (2008).
Solution structure of a conformationally restricted fully active derivative of the human relaxin-like factor.
  Biochemistry, 47, 13308-13317.
PDB codes: 2k6t 2k6u
18332129 Q.X.Hua, S.H.Nakagawa, W.Jia, K.Huang, N.B.Phillips, S.Q.Hu, and M.A.Weiss (2008).
Design of an active ultrastable single-chain insulin analog: synthesis, structure, and therapeutic implications.
  J Biol Chem, 283, 14703-14716.
PDB codes: 2jzq 3bxq
18492668 Z.L.Wan, K.Huang, S.Q.Hu, J.Whittaker, and M.A.Weiss (2008).
The structure of a mutant insulin uncouples receptor binding from protein allostery. An electrostatic block to the TR transition.
  J Biol Chem, 283, 21198-21210.  
17884811 K.Huang, S.J.Chan, Q.X.Hua, Y.C.Chu, R.Y.Wang, B.Klaproth, W.Jia, J.Whittaker, P.De Meyts, S.H.Nakagawa, D.F.Steiner, P.G.Katsoyannis, and M.A.Weiss (2007).
The A-chain of insulin contacts the insert domain of the insulin receptor. Photo-cross-linking and mutagenesis of a diabetes-related crevice.
  J Biol Chem, 282, 35337-35349.
PDB codes: 2jum 2juu 2juv
16627943 J.L.Whittingham, Z.Youshang, L.Záková, E.J.Dodson, J.P.Turkenburg, J.Brange, and G.G.Dodson (2006).
I222 crystal form of despentapeptide (B26-B30) insulin provides new insights into the properties of monomeric insulin.
  Acta Crystallogr D Biol Crystallogr, 62, 505-511.
PDB code: 2ceu
17716170 M.Koch, F.F.Schmid, V.Zoete, and M.Meuwly (2006).
Insulin: a model system for nanomedicine?
  Nanomed, 1, 373-378.  
16894147 M.Lou, T.P.Garrett, N.M.McKern, P.A.Hoyne, V.C.Epa, J.D.Bentley, G.O.Lovrecz, L.J.Cosgrove, M.J.Frenkel, and C.W.Ward (2006).
The first three domains of the insulin receptor differ structurally from the insulin-like growth factor 1 receptor in the regions governing ligand specificity.
  Proc Natl Acad Sci U S A, 103, 12429-12434.
PDB code: 2hr7
16864583 Q.X.Hua, J.P.Mayer, W.Jia, J.Zhang, and M.A.Weiss (2006).
The folding nucleus of the insulin superfamily: a flexible peptide model foreshadows the native state.
  J Biol Chem, 281, 28131-28142.  
16728398 Q.X.Hua, M.Liu, S.Q.Hu, W.Jia, P.Arvan, and M.A.Weiss (2006).
A conserved histidine in insulin is required for the foldability of human proinsulin: structure and function of an ALAB5 analog.
  J Biol Chem, 281, 24889-24899.
PDB code: 2h67
16751187 S.H.Nakagawa, Q.X.Hua, S.Q.Hu, W.Jia, S.Wang, P.G.Katsoyannis, and M.A.Weiss (2006).
Chiral mutagenesis of insulin. Contribution of the B20-B23 beta-turn to activity and stability.
  J Biol Chem, 281, 22386-22396.  
16981237 V.Zoete, and M.Meuwly (2006).
Importance of individual side chains for the stability of a protein fold: computational alanine scanning of the insulin monomer.
  J Comput Chem, 27, 1843-1857.  
15936977 A.Denley, L.J.Cosgrove, G.W.Booker, J.C.Wallace, and B.E.Forbes (2005).
Molecular interactions of the IGF system.
  Cytokine Growth Factor Rev, 16, 421-439.  
15799978 J.Whittaker, and L.Whittaker (2005).
Characterization of the functional insulin binding epitopes of the full-length insulin receptor.
  J Biol Chem, 280, 20932-20936.  
15722444 V.Alexandrov, U.Lehnert, N.Echols, D.Milburn, D.Engelman, and M.Gerstein (2005).
Normal modes for predicting protein motions: a comprehensive database assessment and associated Web tool.
  Protein Sci, 14, 633-643.  
16080143 V.Zoete, M.Meuwly, and M.Karplus (2005).
Study of the insulin dimerization: binding free energy calculations and per-residue free energy decomposition.
  Proteins, 61, 79-93.  
15551269 P.De Meyts (2004).
Insulin and its receptor: structure, function and evolution.
  Bioessays, 26, 1351-1362.  
14988398 Q.X.Hua, and M.A.Weiss (2004).
Mechanism of insulin fibrillation: the structure of insulin under amyloidogenic conditions resembles a protein-folding intermediate.
  J Biol Chem, 279, 21449-21460.
PDB code: 1sf1
12631285 T.Yamazaki, M.Takaoka, E.Katoh, K.Hanada, M.Sakita, K.Sakata, Y.Nishiuchi, and H.Hirano (2003).
A possible physiological function and the tertiary structure of a 4-kDa peptide in legumes.
  Eur J Biochem, 270, 1269-1276.
PDB code: 1ju8
14596591 Z.L.Wan, B.Xu, Y.C.Chu, P.G.Katsoyannis, and M.A.Weiss (2003).
Crystal structure of allo-Ile(A2)-insulin, an inactive chiral analogue: implications for the mechanism of receptor binding.
  Biochemistry, 42, 12770-12783.
PDB codes: 1lw8 1pc1 1q4v
11742127 B.Xu, Q.X.Hua, S.H.Nakagawa, W.Jia, Y.C.Chu, P.G.Katsoyannis, and M.A.Weiss (2002).
A cavity-forming mutation in insulin induces segmental unfolding of a surrounding alpha-helix.
  Protein Sci, 11, 104-116.
PDB code: 1k3m
12360255 P.De Meyts, and J.Whittaker (2002).
Structural biology of insulin and IGF1 receptors: implications for drug design.
  Nat Rev Drug Discov, 1, 769-783.  
12196530 Q.X.Hua, Y.C.Chu, W.Jia, N.F.Phillips, R.Y.Wang, P.G.Katsoyannis, and M.A.Weiss (2002).
Mechanism of insulin chain combination. Asymmetric roles of A-chain alpha-helices in disulfide pairing.
  J Biol Chem, 277, 43443-43453.
PDB code: 1lkq
11500492 J.Whittaker, A.V.Groth, D.C.Mynarcik, L.Pluzek, V.L.Gadsbøll, and L.J.Whittaker (2001).
Alanine scanning mutagenesis of a type 1 insulin-like growth factor receptor ligand binding site.
  J Biol Chem, 276, 43980-43986.  
11064376 L.Nielsen, S.Frokjaer, J.F.Carpenter, and J.Brange (2001).
Studies of the structure of insulin fibrils by Fourier transform infrared (FTIR) spectroscopy and electron microscopy.
  J Pharm Sci, 90, 29-37.  
11517220 M.A.Weiss, Q.X.Hua, W.Jia, S.H.Nakagawa, Y.C.Chu, S.Q.Hu, and P.G.Katsoyannis (2001).
Activities of monomeric insulin analogs at position A8 are uncorrelated with their thermodynamic stabilities.
  J Biol Chem, 276, 40018-40024.  
11447105 W.Zesławski, H.G.Beisel, M.Kamionka, W.Kalus, R.A.Engh, R.Huber, K.Lang, and T.A.Holak (2001).
The interaction of insulin-like growth factor-I with the N-terminal domain of IGFBP-5.
  EMBO J, 20, 3638-3644.
PDB code: 1h59
18763100 Y.Huang, Z.Liang, and Y.Feng (2001).
The relationship between the connecting peptide of recombined single chain insulin and its biological function.
  Sci China C Life Sci, 44, 593-600.  
11258877 Z.S.Qiao, Z.Y.Guo, and Y.M.Feng (2001).
Putative disulfide-forming pathway of porcine insulin precursor during its refolding in vitro.
  Biochemistry, 40, 2662-2668.  
10933788 G.Fullbright, and E.E.Büllesbach (2000).
The receptor binding conformation of bombyxin is induced by alanine(B15).
  Biochemistry, 39, 9718-9724.  
11112528 M.A.Weiss, Q.X.Hua, W.Jia, Y.C.Chu, R.Y.Wang, and P.G.Katsoyannis (2000).
Hierarchical protein "un-design": insulin's intrachain disulfide bridge tethers a recognition alpha-helix.
  Biochemistry, 39, 15429-15440.  
11123908 S.H.Nakagawa, H.S.Tager, and D.F.Steiner (2000).
Mutational analysis of invariant valine B12 in insulin: implications for receptor binding.
  Biochemistry, 39, 15826-15835.  
10448965 E.E.Büllesbach (1999).
Bombyxin exhibits an insulin-like response to modification in the N-terminal region of the A chain.
  J Pept Res, 54, 12-17.  
  10091652 G.Kurapkat, M.Siedentop, H.G.Gattner, M.Hagelstein, D.Brandenburg, J.Grötzinger, and A.Wollmer (1999).
The solution structure of a superpotent B-chain-shortened single-replacement insulin analogue.
  Protein Sci, 8, 499-508.
PDB code: 1bzv
10497806 N.P.wing, and C.J.Cassady (1999).
Effects of cysteic acid groups on the gas-phase reactivity and dissociation of [M + 4H]4+ ions from insulin chain B.
  J Am Soc Mass Spectrom, 10, 928-940.  
10489447 Z.P.Yao, Z.H.Zeng, H.M.Li, Y.Zhang, Y.M.Feng, and D.C.Wang (1999).
Structure of an insulin dimer in an orthorhombic crystal: the structure analysis of a human insulin mutant (B9 Ser-->Glu).
  Acta Crystallogr D Biol Crystallogr, 55, 1524-1532.
PDB code: 1b9e
9148904 C.Kristensen, T.Kjeldsen, F.C.Wiberg, L.Schäffer, M.Hach, S.Havelund, J.Bass, D.F.Steiner, and A.S.Andersen (1997).
Alanine scanning mutagenesis of insulin.
  J Biol Chem, 272, 12978-12983.  
9566117 C.McInnes, and B.D.Sykes (1997).
Growth factor receptors: structure, mechanism, and drug discovery.
  Biopolymers, 43, 339-366.  
8999905 D.C.Mynarcik, P.F.Williams, L.Schaffer, G.Q.Yu, and J.Whittaker (1997).
Analog binding properties of insulin receptor mutants. Identification of amino acids interacting with the COOH terminus of the B-chain of the insulin molecule.
  J Biol Chem, 272, 2077-2081.  
9228034 D.C.Mynarcik, P.F.Williams, L.Schaffer, G.Q.Yu, and J.Whittaker (1997).
Identification of common ligand binding determinants of the insulin and insulin-like growth factor 1 receptors. Insights into mechanisms of ligand binding.
  J Biol Chem, 272, 18650-18655.  
  9070440 G.Kurapkat, E.De Wolf, J.Grötzinger, and A.Wollmer (1997).
Inactive conformation of an insulin despite its wild-type sequence.
  Protein Sci, 6, 580-587.  
9131992 I.Pittman, S.H.Nakagawa, H.S.Tager, and D.F.Steiner (1997).
Maintenance of the B-chain beta-turn in [GlyB24] insulin mutants: a steady-state fluorescence anisotropy study.
  Biochemistry, 36, 3430-3437.  
9035130 N.S.Greenspan (1997).
Reflections on internal images.
  Nat Biotechnol, 15, 123-124.  
9130695 S.Rüdiger, L.Germeroth, J.Schneider-Mergener, and B.Bukau (1997).
Substrate specificity of the DnaK chaperone determined by screening cellulose-bound peptide libraries.
  EMBO J, 16, 1501-1507.  
9235985 X.Chang, A.M.Jorgensen, P.Bardrum, and J.J.Led (1997).
Solution structures of the R6 human insulin hexamer,.
  Biochemistry, 36, 9409-9422.
PDB codes: 1ai0 1aiy
  8931138 E.De Wolf, R.Gill, S.Geddes, J.Pitts, A.Wollmer, and J.Grötzinger (1996).
Solution structure of a mini IGF-1.
  Protein Sci, 5, 2193-2202.
PDB code: 1b9g
7588759 D.R.Hodgson, F.E.May, and B.R.Westley (1995).
Mutations at positions 11 and 60 of insulin-like growth factor 1 reveal differences between its interactions with the type I insulin-like-growth-factor receptor and the insulin receptor.
  Eur J Biochem, 233, 299-309.  
7749917 Q.X.Hua, S.N.Gozani, R.E.Chance, J.A.Hoffmann, B.H.Frank, and M.A.Weiss (1995).
Structure of a protein in a kinetic trap.
  Nat Struct Biol, 2, 129-138.
PDB codes: 1xgl 2hiu
  8011176 A.G.Schepky, A.M.Schmidt, T.Schmidt, P.Schulz-Knappe, and W.G.Forssmann (1994).
Determination of sulfated peptides by differential iodination.
  Biol Chem Hoppe Seyler, 375, 201-203.  
  7527339 H.Terasawa, D.Kohda, H.Hatanaka, K.Nagata, N.Higashihashi, H.Fujiwara, K.Sakano, and F.Inagaki (1994).
Solution structure of human insulin-like growth factor II; recognition sites for receptors and binding proteins.
  EMBO J, 13, 5590-5597.  
7809046 O.Gursky, E.Fontano, B.Bhyravbhatla, and D.L.Caspar (1994).
Stereospecific dihaloalkane binding in a pH-sensitive cavity in cubic insulin crystals.
  Proc Natl Acad Sci U S A, 91, 12388-12392.  
7891290 S.D.Yeo, P.G.Debendetti, S.Y.Patro, and T.M.Przybycien (1994).
Secondary structure characterization of microparticulate insulin powders.
  J Pharm Sci, 83, 1651-1656.  
8421693 Q.X.Hua, S.E.Shoelson, K.Inouye, and M.A.Weiss (1993).
Paradoxical structure and function in a mutant human insulin associated with diabetes mellitus.
  Proc Natl Acad Sci U S A, 90, 582-586.
PDB code: 1hiq
1322139 J.Murray-Rust, A.N.McLeod, T.L.Blundell, and S.P.Wood (1992).
Structure and evolution of insulins: implications for receptor binding.
  Bioessays, 14, 325-331.  
1477273 O.Gursky, J.Badger, Y.Li, and D.L.Caspar (1992).
Conformational changes in cubic insulin crystals in the pH range 7-11.
  Biophys J, 63, 1210-1220.
PDB codes: 1aph 1bph 1cph 1dph
1369391 P.R.Young (1992).
Protein hormones and their receptors.
  Curr Opin Biotechnol, 3, 408-421.  
1549601 Q.X.Hua, M.Kochoyan, and M.A.Weiss (1992).
Structure and dynamics of des-pentapeptide-insulin in solution: the molten-globule hypothesis.
  Proc Natl Acad Sci U S A, 89, 2379-2383.
PDB code: 1his
1511237 Y.Liu, D.Zhao, R.Altman, and O.Jardetzky (1992).
A systematic comparison of three structure determination methods from NMR data: dependence upon quality and quantity of data.
  J Biomol NMR, 2, 373-388.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB codes are shown on the right.