PDBsum entry 2hh4

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Hormone/growth factor PDB id
Protein chains
21 a.a.
30 a.a. *
* Residue conservation analysis
PDB id:
Name: Hormone/growth factor
Title: Nmr structure of human insulin mutant gly-b8-d-ser, his-b10- asp pro-b28-lys, lys-b29-pro, 20 structures
Structure: Insulin a chain. Chain: a. Engineered: yes. Insulin b chain. Chain: b. Engineered: yes. Mutation: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: ins. Expressed in: escherichia coli. Expression_system_taxid: 562. Expression_system_taxid: 562
NMR struc: 20 models
Authors: Q.X.Hua,S.Nakagawa,S.Q.Hu,W.Jia,M.A.Weiss
Key ref:
Q.X.Hua et al. (2006). Toward the active conformation of insulin: stereospecific modulation of a structural switch in the B chain. J Biol Chem, 281, 24900-24909. PubMed id: 16762918 DOI: 10.1074/jbc.M602691200
27-Jun-06     Release date:   18-Jul-06    
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)

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


DOI no: 10.1074/jbc.M602691200 J Biol Chem 281:24900-24909 (2006)
PubMed id: 16762918  
Toward the active conformation of insulin: stereospecific modulation of a structural switch in the B chain.
Q.X.Hua, S.Nakagawa, S.Q.Hu, W.Jia, S.Wang, M.A.Weiss.
How insulin binds to the insulin receptor has long been a subject of speculation. Although the structure of the free hormone has been extensively characterized, a variety of evidence suggests that a conformational change occurs upon receptor binding. Here, we employ chiral mutagenesis, comparison of corresponding d and l amino acid substitutions, to investigate a possible switch in the B-chain. To investigate the interrelation of structure, function, and stability, isomeric analogs have been synthesized in which an invariant glycine in a beta-turn (Gly(B8)) is replaced by d- or l-Ser. The d substitution enhances stability (DeltaDeltaG(u) 0.9 kcal/mol) but impairs receptor binding by 100-fold; by contrast, the l substitution markedly impairs stability (DeltaDeltaG(u) -3.0 kcal/mol) with only 2-fold reduction in receptor binding. Although the isomeric structures each retain a native-like overall fold, the l-Ser(B8) analog exhibits fewer helix-related and long range nuclear Overhauser effects than does the d-Ser(B8) analog or native monomer. Evidence for enhanced conformational fluctuations in the unstable analog is provided by its attenuated CD spectrum. The inverse relationship between stereospecific stabilization and receptor binding strongly suggests that the B7-B10 beta-turn changes conformation on receptor binding.
  Selected figure(s)  
Figure 3.
FIGURE 3. Two-dimensional NMR identification of D- and L-Ser^B8 spin systems and diagonal plot of interresidue NOES. TOCSY spectra of D-Ser^B8-DKP-insulin (A) and L-Ser^B8-DKP-insulin (B) in the region containing the AMX spin system of respective B8 side chains. Spectra (mixing times 55 ms) were observed in D[2]O at 32 °C and pD 7.6 (direct meter reading). C and D, diagonal plot of D-Ser^B8-DKP-insulin (C) and L-Ser^B8-DKP-insulin (D) shown inter-residue NOEs, respectively. NOEs between side chains are shown at lower right (open boxes); NOEs between main chain protons or between main chain and side chains are shown at upper left (filled boxes). Red squares in C indicate NOEs consistent with T-state crystal structures but unobserved in the spectrum of L-Ser^B8-DKP-insulin. In C, NOEs a-c indicate (a) set of contacts between the Ile^A10 side chain and main chain atoms of Asn^B3, Gln^B4, and His^B5; (b) contact between the side chains of His^B5 and Thr^A8; and (c) the set of contacts between the Leu^B6 side chain and H[ ]of Leu^B11 across the -turn. In D, red boxes indicate NOEs present in the L analog but not in the D analog. Respective green and red boxes d and e indicate (d) Cys^A11 side chain to H[ ]of Cys^A6; and (e) contact between H[ ]of Tyr^A19 and meta resonance of Phe^B24.
Figure 5.
FIGURE 5. DG/RMD ensembles. Front and back views of D-Ser^B8-DKP-insulin (A) and L-Ser^B8-DKP-insulin (B). In each case the A-chain is shown in gray and B-chain in blue. Left and right images are related by a rotation of 180° about the vertical axis. The D- and L-Ser^B8 side chains are shown in green and purple, respectively (arrowheads). Ribbons indicate the mean structure of DKP-insulin. Arrow indicates the region of decreased precision in the D analog; asterisk indicates the region of decreased precision in the L analog. Structures were aligned according to the main atom chains of residues A2-A7, A13-A19, and B9-B23.
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2006, 281, 24900-24909) copyright 2006.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
20948967 M.Liu, L.Haataja, J.Wright, N.P.Wickramasinghe, Q.X.Hua, N.F.Phillips, F.Barbetti, M.A.Weiss, and P.Arvan (2010).
Mutant INS-gene induced diabetes of youth: proinsulin cysteine residues impose dominant-negative inhibition on wild-type proinsulin transport.
  PLoS One, 5, e13333.  
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.  
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.  
19395706 M.A.Weiss (2009).
Proinsulin and the genetics of diabetes mellitus.
  J Biol Chem, 284, 19159-19163.  
19850922 M.Liu, Z.L.Wan, Y.C.Chu, H.Aladdin, B.Klaproth, M.Choquette, Q.X.Hua, R.B.Mackin, J.S.Rao, P.De Meyts, P.G.Katsoyannis, P.Arvan, and M.A.Weiss (2009).
Crystal structure of a "nonfoldable" insulin: impaired folding efficiency despite native activity.
  J Biol Chem, 284, 35259-35272.
PDB code: 3gky
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
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.  
17855560 J.Støy, E.L.Edghill, S.E.Flanagan, H.Ye, V.P.Paz, A.Pluzhnikov, J.E.Below, M.G.Hayes, N.J.Cox, G.M.Lipkind, R.B.Lipton, S.A.Greeley, A.M.Patch, S.Ellard, D.F.Steiner, A.T.Hattersley, L.H.Philipson, and G.I.Bell (2007).
Insulin gene mutations as a cause of permanent neonatal diabetes.
  Proc Natl Acad Sci U S A, 104, 15040-15044.  
17436342 L.Záková, D.Zyka, J.Jezek, I.Hanclová, M.Sanda, A.M.Brzozowski, and J.Jirácek (2007).
The use of Fmoc-Lys(Pac)-OH and penicillin G acylase in the preparation of novel semisynthetic insulin analogs.
  J Pept Sci, 13, 334-341.  
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.