PDBsum entry 1lph

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protein ligands metals Protein-protein interface(s) links
Hormone PDB id
Protein chains
21 a.a.
30 a.a. *
_ZN ×2
Waters ×59
* Residue conservation analysis
PDB id:
Name: Hormone
Title: Lys(b28)pro(b29)-human insulin
Structure: Insulin. Chain: a, c. Synonym: lys(b28)pro(b29)-human insulin. Engineered: yes. Mutation: yes. Insulin. Chain: b, d. Synonym: lys(b28)pro(b29)-human insulin. Engineered: yes.
Source: Homo sapiens. Human. Organism_taxid: 9606. Organism_taxid: 9606
Biol. unit: Dodecamer (from PDB file)
2.30Å     R-factor:   0.161    
Authors: E.Ciszak,J.M.Beals,B.H.Frank,J.C.Baker,N.D.Carter,G.D.Smith
Key ref:
E.Ciszak et al. (1995). Role of C-terminal B-chain residues in insulin assembly: the structure of hexameric LysB28ProB29-human insulin. Structure, 3, 615-622. PubMed id: 8590022 DOI: 10.1016/S0969-2126(01)00195-2
19-Apr-95     Release date:   20-Jun-96    
Go to PROCHECK summary

Protein chains
Pfam   ArchSchema ?
P01308  (INS_HUMAN) -  Insulin
110 a.a.
21 a.a.
Protein chains
Pfam   ArchSchema ?
P01308  (INS_HUMAN) -  Insulin
110 a.a.
30 a.a.*
Key:    PfamA domain  Secondary structure
* PDB and UniProt seqs differ at 2 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: 10.1016/S0969-2126(01)00195-2 Structure 3:615-622 (1995)
PubMed id: 8590022  
Role of C-terminal B-chain residues in insulin assembly: the structure of hexameric LysB28ProB29-human insulin.
E.Ciszak, J.M.Beals, B.H.Frank, J.C.Baker, N.D.Carter, G.D.Smith.
BACKGROUND: LysB28ProB29-human insulin (Humalog), a fully potent insulin analog in which the prolyl, lysyl sequence at the C-terminal end of the B-chain is inverted, exhibits a decreased association of monomers to dimers leading to rapid in vivo absorption. This provides important benefits for the insulin-requiring diabetic. In spite of its monomeric nature, LysB28ProB29-human insulin can exist as a discrete hexameric structure in the presence of both zinc and phenol. Studies of the crystal structure of LysB28ProB29-human insulin in a hexameric complex were initiated to gain a molecular understanding of the effect of the sequence inversion on the analog's self-association properties and, consequently, its in vivo efficacy. RESULTS: Under the conditions reported, LysB28ProB29-human insulin crystallized as a T3Rf3 hexamer that is isomorphous with the uncomplexed T3Rf3 native human insulin hexamer previously known as '4Zn insulin'. The three-dimensional structure of the T3Rf3 hexamer was determined by X-ray crystallographic methods to a resolution of 2.3 A. The prolyl, lysyl sequence inversion leads to local conformational changes at the C termini of the B-chains which eliminate two critical hydrophobic interactions and weaken two terminal beta-sheet hydrogen bonds that stabilize the dimer. CONCLUSIONS: The loss of these native dimer interactions weakens the hexameric LysB28ProB29-human insulin complex formed in the presence of phenolic ligands. Thus, it is hypothesized that the diffusion of the phenolic ligands from the site of injection results in the dissociation of hexamers directly to monomers, thereby maintaining the rapid time-action of the monomeric analog in spite of the hexameric conformation in therapeutic formulations.
  Selected figure(s)  
Figure 6.
Figure 6. A superposition of the sequence Tyr^B26.1–Thr^B30.1 in human insulin (red) as compared with the same fragment in Lys^B28Pro^B29-human insulin (colored according to atom type). Figure 6. A superposition of the sequence Tyr^B26.1–Thr^B30.1 in human insulin (red) as compared with the same fragment in Lys^B28Pro^B29-human insulin (colored according to atom type).
Figure 8.
Figure 8. 2F[o]−F[c] electron density maps contoured at 1 σ in the vicinity of the B-chain C-terminal residues of (a) the T-state monomer and (b) the R^f-state monomer. Figure 8. 2F[o]−F[c] electron density maps contoured at 1 σ in the vicinity of the B-chain C-terminal residues of (a) the T-state monomer and (b) the R^f-state monomer.
  The above figures are reprinted by permission from Cell Press: Structure (1995, 3, 615-622) copyright 1995.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
20070212 Y.B.Chen, J.Li, Y.Qi, X.Miao, Y.Zhou, D.Ren, and G.Z.Guo (2010).
The effects of electromagnetic pulses (EMP) on the bioactivity of insulin and a preliminary study of mechanism.
  Int J Radiat Biol, 86, 22-26.  
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
18260111 W.Bocian, J.Sitkowski, A.Tarnowska, E.Bednarek, R.Kawecki, W.Koźmiński, and L.Kozerski (2008).
Direct insight into insulin aggregation by 2D NMR complemented by PFGSE NMR.
  Proteins, 71, 1057-1065.  
18040865 W.Bocian, J.Sitkowski, E.Bednarek, A.Tarnowska, R.Kawecki, and L.Kozerski (2008).
Structure of human insulin monomer in water/acetonitrile solution.
  J Biomol NMR, 40, 55-64.
PDB code: 2jv1
18491415 W.Bocian, P.Borowicz, J.Mikołajczyk, J.Sitkowski, A.Tarnowska, E.Bednarek, T.Głabski, B.Tejchman-Małecka, M.Bogiel, and L.Kozerski (2008).
NMR structure of biosynthetic engineered human insulin monomer B31(Lys)-B32(Arg) in water/acetonitrile solution. Comparison with the solution structure of native human insulin monomer.
  Biopolymers, 89, 820-830.
PDB code: 2rn5
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.  
17335298 D.Simpson, P.L.McCormack, G.M.Keating, and K.A.Lyseng-Williamson (2007).
Insulin lispro: a review of its use in the management of diabetes mellitus.
  Drugs, 67, 407-434.  
17410596 J.P.Mayer, F.Zhang, and R.D.DiMarchi (2007).
Insulin structure and function.
  Biopolymers, 88, 687-713.  
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
17316105 R.H.Becker (2007).
Insulin glulisine complementing basal insulins: a review of structure and activity.
  Diabetes Technol Ther, 9, 109-121.  
  16820682 E.K.Millers, P.P.Masci, M.F.Lavin, Jersey, and L.W.Guddat (2006).
Crystallization and preliminary X-ray analysis of a Kunitz-type inhibitor, textilinin-1 from Pseudonaja textilis textilis.
  Acta Crystallogr Sect F Struct Biol Cryst Commun, 62, 642-645.  
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
16239724 G.D.Smith, W.A.Pangborn, and R.H.Blessing (2005).
The structure of T6 bovine insulin.
  Acta Crystallogr D Biol Crystallogr, 61, 1476-1482.
PDB code: 2a3g
15580495 G.Walsh (2005).
Therapeutic insulins and their large-scale manufacture.
  Appl Microbiol Biotechnol, 67, 151-159.  
16254391 L.Bromberg, J.Rashba-Step, and T.Scott (2005).
Insulin particle formation in supersaturated aqueous solutions of poly(ethylene glycol).
  Biophys J, 89, 3424-3433.  
15776817 S.K.Rastogi, and J.Singh (2005).
Effect of chemical penetration enhancer and iontophoresis on the in vitro percutaneous absorption enhancement of insulin through porcine epidermis.
  Pharm Dev Technol, 10, 97.  
12446709 B.Y.Zhang, M.Liu, and P.Arvan (2003).
Behavior in the eukaryotic secretory pathway of insulin-containing fusion proteins and single-chain insulins bearing various B-chain mutations.
  J Biol Chem, 278, 3687-3693.  
12764141 C.C.Yip, and P.Ottensmeyer (2003).
Three-dimensional structural interactions of insulin and its receptor.
  J Biol Chem, 278, 27329-27332.  
12876340 G.D.Smith, and R.H.Blessing (2003).
Lessons from an aged, dried crystal of T(6) human insulin.
  Acta Crystallogr D Biol Crystallogr, 59, 1384-1394.
PDB codes: 1os3 1os4
12930990 H.B.Olsen, M.R.Leuenberger-Fisher, W.Kadima, D.Borchardt, N.C.Kaarsholm, and M.F.Dunn (2003).
Structural signatures of the complex formed between 3-nitro-4-hydroxybenzoate and the Zn(II)-substituted R(6) insulin hexamer.
  Protein Sci, 12, 1902-1913.  
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
11745120 D.Ferrari, J.R.Diers, D.F.Bocian, N.C.Kaarsholm, and M.F.Dunn (2001).
Raman signatures of ligand binding and allosteric conformation change in hexameric insulin.
  Biopolymers, 62, 249-260.  
11468392 G.D.Smith, W.A.Pangborn, and R.H.Blessing (2001).
Phase changes in T(3)R(3)(f) human insulin: temperature or pressure induced?
  Acta Crystallogr D Biol Crystallogr, 57, 1091-1100.
PDB codes: 1g7a 1g7b
11343787 J.Ye, W.Chang, and D.Liang (2001).
Crystal structure of destripeptide (B28-B30) insulin: implications for insulin dissociation.
  Biochim Biophys Acta, 1547, 18-25.
PDB code: 1htv
10620310 C.M.Yip, M.L.Brader, B.H.Frank, M.R.DeFelippis, and M.D.Ward (2000).
Structural studies of a crystalline insulin analog complex with protamine by atomic force microscopy.
  Biophys J, 78, 466-473.  
11092919 G.D.Smith, E.Ciszak, L.A.Magrum, W.A.Pangborn, and R.H.Blessing (2000).
R6 hexameric insulin complexed with m-cresol or resorcinol.
  Acta Crystallogr D Biol Crystallogr, 56, 1541-1548.
PDB codes: 1ev3 1ev6 1evr
10981628 J.McCafferty, and D.R.Glover (2000).
Engineering therapeutic proteins.
  Curr Opin Struct Biol, 10, 417-420.  
10746744 J.Shneine, M.Voswinkel, M.Federwisch, and A.Wollmer (2000).
Enhancing the T-->R transition of insulin by helix-promoting sequence modifications at the N-terminal B-chain.
  Biol Chem, 381, 127-133.  
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.  
10397800 H.Berchtold, and R.Hilgenfeld (1999).
Binding of phenol to R6 insulin hexamers.
  Biopolymers, 51, 165-172.  
10837704 J.Brange, and A.Vølund (1999).
Insulin analogs with improved pharmacokinetic profiles.
  Adv Drug Deliv Rev, 35, 307-335.  
10479347 J.P.Richards, M.P.Stickelmeyer, B.H.Frank, S.Pye, M.Barbeau, J.Radziuk, G.D.Smith, and M.R.DeFelippis (1999).
Preparation of a microcrystalline suspension formulation of Lys(B28)Pro(B29)-human insulin with ultralente properties.
  J Pharm Sci, 88, 861-867.  
9591647 C.M.Yip, M.L.Brader, M.R.DeFelippis, and M.D.Ward (1998).
Atomic force microscopy of crystalline insulins: the influence of sequence variation on crystallization and interfacial structure.
  Biophys J, 74, 2199-2209.  
9708987 J.L.Whittingham, D.J.Edwards, A.A.Antson, J.M.Clarkson, and G.G.Dodson (1998).
Interactions of phenol and m-cresol in the insulin hexamer, and their effect on the association properties of B28 pro --> Asp insulin analogues.
  Biochemistry, 37, 11516-11523.
PDB codes: 1zeg 1zeh 1zei
9519149 M.R.DeFelippis, D.L.Bakaysa, M.A.Bell, M.A.Heady, S.Li, S.Pye, K.M.Youngman, J.Radziuk, and B.H.Frank (1998).
Preparation and characterization of a cocrystalline suspension of [LysB28,ProB29]-human insulin analogue.
  J Pharm Sci, 87, 170-176.  
8988131 A.H.Barnett, and D.R.Owens (1997).
Insulin analogues.
  Lancet, 349, 47-51.  
9062110 J.L.Whittingham, S.Havelund, and I.Jonassen (1997).
Crystal structure of a prolonged-acting insulin with albumin-binding properties.
  Biochemistry, 36, 2826-2831.
PDB code: 1xda
9153424 S.Rahuel-Clermont, C.A.French, N.C.Kaarsholm, M.F.Dunn, and C.I.Chou (1997).
Mechanisms of stabilization of the insulin hexamer through allosteric ligand interactions.
  Biochemistry, 36, 5837-5845.  
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
  8976561 D.L.Bakaysa, J.Radziuk, H.A.Havel, M.L.Brader, S.Li, S.W.Dodd, J.M.Beals, A.H.Pekar, and D.N.Brems (1996).
Physicochemical basis for the rapid time-action of LysB28ProB29-insulin: dissociation of a protein-ligand complex.
  Protein Sci, 5, 2521-2531.  
  8844841 G.D.Smith, E.Ciszak, and W.Pangborn (1996).
A novel complex of a phenolic derivative with insulin: structural features related to the T-->R transition.
  Protein Sci, 5, 1502-1511.
PDB code: 1ben
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.