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PDBsum entry 1guj

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protein ligands Protein-protein interface(s) links
Hormone PDB id
1guj
Jmol
Contents
Protein chains
21 a.a.
30 a.a. *
Ligands
SO4 ×2
Waters ×127
* Residue conservation analysis
PDB id:
1guj
Name: Hormone
Title: Insulin at ph 2: structural analysis of the conditions promoting insulin fibre formation.
Structure: Insulin. Chain: a, c. Engineered: yes. Insulin. Chain: b, d. Engineered: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Expressed in: saccharomyces cerevisiae. Expression_system_taxid: 4932. Expression_system_taxid: 4932
Biol. unit: Hetero-Dimer (from PDB file)
Resolution:
1.62Å     R-factor:   0.172     R-free:   0.207
Authors: J.L.Whittingham,D.J.Scott,K.Chance,A.Wilson,J.Finch, J.Brange,G.G.Dodson
Key ref:
J.L.Whittingham et al. (2002). Insulin at pH 2: structural analysis of the conditions promoting insulin fibre formation. J Mol Biol, 318, 479-490. PubMed id: 12051853 DOI: 10.1016/S0022-2836(02)00021-9
Date:
28-Jan-02     Release date:   08-Mar-02    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P01308  (INS_HUMAN) -  Insulin
Seq:
Struc:
110 a.a.
21 a.a.
Protein chains
Pfam   ArchSchema ?
P01308  (INS_HUMAN) -  Insulin
Seq:
Struc:
110 a.a.
30 a.a.
Key:    PfamA domain  Secondary structure

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

 

 
DOI no: 10.1016/S0022-2836(02)00021-9 J Mol Biol 318:479-490 (2002)
PubMed id: 12051853  
 
 
Insulin at pH 2: structural analysis of the conditions promoting insulin fibre formation.
J.L.Whittingham, D.J.Scott, K.Chance, A.Wilson, J.Finch, J.Brange, G.Guy Dodson.
 
  ABSTRACT  
 
When insulin solutions are subjected to acid, heat and agitation, the normal pattern of insulin assembly (dimers-->tetramers-->hexamers) is disrupted; the molecule undergoes conformational changes allowing it to follow an alternative aggregation pathway (via a monomeric species) leading to the formation of insoluble amyloid fibres. To investigate the effect of acid pH on the conformation and aggregation state of the protein, the crystal structure of human insulin at pH 2.1 has been determined to 1.6 A resolution. The structure reveals that the native fold is maintained at low pH, and that the molecule is still capable of forming dimers similar to those found in hexameric insulin structures at higher pH. Sulphate ions are incorporated into the molecule and the crystal lattice where they neutralise positive charges on the protein, stabilising its structure and facilitating crystallisation. The sulphate interactions are associated with local deformations in the protein, which may indicate that the structure is more plastic at low pH. Transmission electron microscopy analysis of insulin fibres reveals that the appearance of the fibres is greatly influenced by the type of acid employed. Sulphuric acid produces distinctive highly bunched, truncated fibres, suggesting that the sulphate ions have a sophisticated role to play in fibre formation, rather as they do in the crystal structure. Analytical ultracentrifugation studies show that in the absence of heating, insulin is predominantly dimeric in mineral acids, whereas in acetic acid the equilibrium is shifted towards the monomer. Hence, the effect of acid on the aggregation state of insulin is also complex. These results suggest that acid conditions increase the susceptibility of the molecule to conformational change and dissociation, and enhance the rate of fibrillation by providing a charged environment in which the attractive forces between the protein molecules is increased.
 
  Selected figure(s)  
 
Figure 2.
Figure 2. Negative staining transmission electron micrographs of human insulin fibres prepared under a variety of conditions. Unless otherwise stated, all solutions contained 5 mg/ml human insulin. The conditions used were: (a) 0.004 M H[2]SO[4] (pH 2.1), 70 °C; (b) 0.01 M HCl (pH 2.0), 70 °C; (c) H[3]PO[4] (pH 2.0), 90 °C; (d) 0.5 M citric acid (pH 1.9), 90 °C; (e) 20 mg/ml bovine insulin in 8.3 M acetic acid (pH 1.6), 37 °C; (f) 20% acetic acid (pH 2.0) and 0.004 M Na[2]SO[4], 90 °C.
Figure 3.
Figure 3. Stereo view illustrations showing comparisons of the pH 2 insulin dimer (green) with the dimer of (a) the T[6] insulin hexamer (blue),[20.] and (b) the B9 Ser->Glu mutant insulin dimer (red). [9.] Only C^a atoms and the sulphate ions (ball and stick representation in the low pH structure) are shown. The dimers were overlapped using an alignment on residues B9-B19 and D9-D19, which constitute the B chain a-helices in each dimer. This Figure was made using MOLSCRIPT.[36.]
 
  The above figures are reprinted by permission from Elsevier: J Mol Biol (2002, 318, 479-490) copyright 2002.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
21485029 J.K.Raynes, F.G.Pearce, S.J.Meade, and J.A.Gerrard (2011).
Immobilization of organophosphate hydrolase on an amyloid fibril nanoscaffold: Towards bioremediation and chemical detoxification.
  Biotechnol Prog, 27, 360-367.  
20202737 A.K.Attri, C.Fernández, and A.P.Minton (2010).
pH-dependent self-association of zinc-free insulin characterized by concentration-gradient static light scattering.
  Biophys Chem, 148, 28-33.  
20407709 F.Evers, C.Reichhart, R.Steitz, M.Tolan, and C.Czeslik (2010).
Probing adsorption and aggregation of insulin at a poly(acrylic acid) brush.
  Phys Chem Chem Phys, 12, 4375-4382.  
18655073 A.Nayak, M.Sorci, S.Krueger, and G.Belfort (2009).
A universal pathway for amyloid nucleus and precursor formation for insulin.
  Proteins, 74, 556-565.  
19186147 C.Jeworrek, O.Hollmann, R.Steitz, R.Winter, and C.Czeslik (2009).
Interaction of IAPP and insulin with model interfaces studied using neutron reflectometry.
  Biophys J, 96, 1115-1123.  
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.  
19381627 L.M.Cortez, R.N.Farías, and R.N.Chehín (2009).
Protective effect of 3,5,3'-triiodothyroacetic and 3,5,3',5'-tetraiodothyroacetic acids on serum albumin fibrillation.
  Eur Biophys J, 38, 857-863.  
18452331 A.Top, K.L.Kiick, and C.J.Roberts (2008).
Modulation of self-association and subsequent fibril formation in an alanine-rich helical polypeptide.
  Biomacromolecules, 9, 1595-1603.  
18469080 S.T.Henriques, L.K.Pattenden, M.I.Aguilar, and M.A.Castanho (2008).
PrP(106-126) does not interact with membranes under physiological conditions.
  Biophys J, 95, 1877-1889.  
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.  
17472440 B.Vestergaard, M.Groenning, M.Roessle, J.S.Kastrup, M.van de Weert, J.M.Flink, S.Frokjaer, M.Gajhede, and D.I.Svergun (2007).
A helical structural nucleus is the primary elongating unit of insulin amyloid fibrils.
  PLoS Biol, 5, e134.  
17678264 N.Javid, K.Vogtt, C.Krywka, M.Tolan, and R.Winter (2007).
Capturing the interaction potential of amyloidogenic proteins.
  Phys Rev Lett, 99, 028101.  
17279334 R.Sood, Y.Domanov, and P.K.Kinnunen (2007).
Fluorescent temporin B derivative and its binding to liposomes.
  J Fluoresc, 17, 223-234.  
16239333 A.Podestà, G.Tiana, P.Milani, and M.Manno (2006).
Early events in insulin fibrillization studied by time-lapse atomic force microscopy.
  Biophys J, 90, 589-597.  
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
16537488 M.I.Ivanova, M.J.Thompson, and D.Eisenberg (2006).
A systematic screen of beta(2)-microglobulin and insulin for amyloid-like segments.
  Proc Natl Acad Sci U S A, 103, 4079-4082.  
16272434 R.F.Pasternack, E.J.Gibbs, S.Sibley, L.Woodard, P.Hutchinson, J.Genereux, and K.Kristian (2006).
Formation kinetics of insulin-based amyloid gels and the effect of added metalloporphyrins.
  Biophys J, 90, 1033-1042.  
16246845 A.Ahmad, V.N.Uversky, D.Hong, and A.L.Fink (2005).
Early events in the fibrillation of monomeric insulin.
  J Biol Chem, 280, 42669-42675.  
16322584 F.Librizzi, and C.Rischel (2005).
The kinetic behavior of insulin fibrillation is determined by heterogeneous nucleation pathways.
  Protein Sci, 14, 3129-3134.  
16239223 K.Huang, J.Dong, N.B.Phillips, P.R.Carey, and M.A.Weiss (2005).
Proinsulin is refractory to protein fibrillation: topological protection of a precursor protein from cross-beta assembly.
  J Biol Chem, 280, 42345-42355.  
15574704 R.Jansen, W.Dzwolak, and R.Winter (2005).
Amyloidogenic self-assembly of insulin aggregates probed by high resolution atomic force microscopy.
  Biophys J, 88, 1344-1353.  
14736893 A.Ahmad, I.S.Millett, S.Doniach, V.N.Uversky, and A.L.Fink (2004).
Stimulation of insulin fibrillation by urea-induced intermediates.
  J Biol Chem, 279, 14999-15013.  
15295111 A.Arora, C.Ha, and C.B.Park (2004).
Insulin amyloid fibrillation at above 100 degrees C: new insights into protein folding under extreme temperatures.
  Protein Sci, 13, 2429-2436.  
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
15169954 W.Dzwolak, V.Smirnovas, R.Jansen, and R.Winter (2004).
Insulin forms amyloid in a strain-dependent manner: an FT-IR spectroscopic study.
  Protein Sci, 13, 1927-1932.  
12499373 W.B.Stine, K.N.Dahlgren, G.A.Krafft, and M.J.LaDu (2003).
In vitro characterization of conditions for amyloid-beta peptide oligomerization and fibrillogenesis.
  J Biol Chem, 278, 11612-11622.  
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.