PDBsum entry 2juu

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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: Allo-thra3 dkp-insulin
Structure: Insulin a chain. Chain: a. Engineered: yes. Mutation: yes. Insulin b chain. Chain: b. Engineered: yes. Mutation: yes
Source: Synthetic: yes. Synthetic: yes
NMR struc: 15 models
Authors: K.Huang,S.Chan,Q.Hua,Y.Chu,R.Wang,B.Klaproth,W.Jia, J.Whittaker,P.De Meyts,S.H.Nakagawa,D.F.Steiner, P.G.Katsoyannis,M.A.Weiss
Key ref:
K.Huang et al. (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. PubMed id: 17884811 DOI: 10.1074/jbc.M705996200
03-Sep-07     Release date:   16-Oct-07    
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 4 residue positions (black crosses)


DOI no: 10.1074/jbc.M705996200 J Biol Chem 282:35337-35349 (2007)
PubMed id: 17884811  
The A-chain of insulin contacts the insert domain of the insulin receptor. Photo-cross-linking and mutagenesis of a diabetes-related crevice.
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, M.A.Weiss.
The contribution of the insulin A-chain to receptor binding is investigated by photo-cross-linking and nonstandard mutagenesis. Studies focus on the role of Val(A3), which projects within a crevice between the A- and B-chains. Engineered receptor alpha-subunits containing specific protease sites ("midi-receptors") are employed to map the site of photo-cross-linking by an analog containing a photoactivable A3 side chain (para-azido-Phe (Pap)). The probe cross-links to a C-terminal peptide (residues 703-719 of the receptor A isoform, KTFEDYLHNVVFVPRPS) containing side chains critical for hormone binding (underlined); the corresponding segment of the holoreceptor was shown previously to cross-link to a Pap(B25)-insulin analog. Because Pap is larger than Val and so may protrude beyond the A3-associated crevice, we investigated analogs containing A3 substitutions comparable in size to Val as follows: Thr, allo-Thr, and alpha-aminobutyric acid (Aba). Substitutions were introduced within an engineered monomer. Whereas previous studies of smaller substitutions (Gly(A3) and Ser(A3)) encountered nonlocal conformational perturbations, NMR structures of the present analogs are similar to wild-type insulin; the variant side chains are accommodated within a native-like crevice with minimal distortion. Receptor binding activities of Aba(A3) and allo-Thr(A3) analogs are reduced at least 10-fold; the activity of Thr(A3)-DKP-insulin is reduced 5-fold. The hormone-receptor interface is presumably destabilized either by a packing defect (Aba(A3)) or by altered polarity (allo-Thr(A3) and Thr(A3)). Our results provide evidence that Val(A3), a site of mutation causing diabetes mellitus, contacts the insert domain-derived tail of the alpha-subunit in a hormone-receptor complex.
  Selected figure(s)  
Figure 2.
FIGURE 2. Stereochemical configurations of variant amino acids. Aba, an unbranched nonstandard residue, has one -methyl group. Thr and allo-Thr (stereoisomers related by inversion of chirality at the β-carbon) have one methyl group and one β-hydroxyl group.
Figure 8.
FIGURE 8. Crystal structure of receptor ectodomain and possible position of CT segment. A, ribbon model of component protomer (PDB code, 2DTG). Individual domains L1, CR, L2, FnIII-1, FnIII-2, and FnIII-3 are shown in gray, black, blue, red, purple, and dark blue, respectively. Missing or discontinuous electron density (residues 655-755; IR isoform A) is depicted in schematic form by red dashed lines (not intended to represent actual conformation). B, crystal structure of inverted V ectodomain dimer. C, L1 domain (gray ribbon) in relation to unassigned and discontinuous electron density (green), potentially from CT; density may represent 20 residues of insert domain. Residues in L1 critical to hormone binding (as inferred from Ala-scanning mutagenesis (93) are shown as red stick. C was kindly provided by C. W. Ward.
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2007, 282, 35337-35349) copyright 2007.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
20348418 B.J.Smith, K.Huang, G.Kong, S.J.Chan, S.Nakagawa, J.G.Menting, S.Q.Hu, J.Whittaker, D.F.Steiner, P.G.Katsoyannis, C.W.Ward, M.A.Weiss, and M.C.Lawrence (2010).
Structural resolution of a tandem hormone-binding element in the insulin receptor and its implications for design of peptide agonists.
  Proc Natl Acad Sci U S A, 107, 6771-6776.
PDB code: 3loh
20862723 R.A.Serwa, J.M.Swiecicki, D.Homann, and C.P.Hackenberger (2010).
Phosphoramidate-peptide synthesis by solution- and solid-phase Staudinger-phosphite reactions.
  J Pept Sci, 16, 563-567.  
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.  
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.  
19618407 G.Le Flem, J.Pecher, V.Le Flem-Bonhomme, A.Withdrawn, J.Rochette, J.P.Pujol, and P.Bogdanowicz (2009).
Human insulin A-chain peptide analog(s) with in vitro biological activity.
  Cell Biochem Funct, 27, 370-377.  
19321446 M.Manolopoulou, Q.Guo, E.Malito, A.B.Schilling, and W.J.Tang (2009).
Molecular basis of catalytic chamber-assisted unfolding and cleavage of human insulin by human insulin-degrading enzyme.
  J Biol Chem, 284, 14177-14188.
PDB codes: 2wby 2wc0
19773552 M.Zhao, Z.L.Wan, L.Whittaker, B.Xu, N.B.Phillips, P.G.Katsoyannis, F.Ismail-Beigi, J.Whittaker, and M.A.Weiss (2009).
Design of an insulin analog with enhanced receptor binding selectivity: rationale, structure, and therapeutic implications.
  J Biol Chem, 284, 32178-32187.
PDB code: 3fq9
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
18640841 P.De Meyts (2008).
The insulin receptor: a prototype for dimeric, allosteric membrane receptors?
  Trends Biochem Sci, 33, 376-384.  
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.  
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 code is shown on the right.