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PDBsum entry 1hui

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protein Protein-protein interface(s) links
Hormone PDB id
1hui
Jmol
Contents
Protein chains
21 a.a.
29 a.a.
PDB id:
1hui
Name: Hormone
Title: Insulin mutant (b1, b10, b16, b27)glu, des-b30, nmr, 25 structures
Structure: Insulin. Chain: a. Engineered: yes. Mutation: yes. Insulin. Chain: b. Engineered: yes. Mutation: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Organ: pancreas. Expressed in: saccharomyces cerevisiae. Expression_system_taxid: 4932. Expression_system_taxid: 4932
NMR struc: 25 models
Authors: H.B.Olsen,S.Ludvigsen,N.C.Kaarsholm
Key ref:
H.B.Olsen et al. (1996). Solution structure of an engineered insulin monomer at neutral pH. Biochemistry, 35, 8836-8845. PubMed id: 8688419 DOI: 10.1021/bi960292+
Date:
29-Mar-96     Release date:   12-Mar-97    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P01308  (INS_HUMAN) -  Insulin
Seq:
Struc:
110 a.a.
21 a.a.
Protein chain
Pfam   ArchSchema ?
P01308  (INS_HUMAN) -  Insulin
Seq:
Struc:
110 a.a.
29 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: 10.1021/bi960292+ Biochemistry 35:8836-8845 (1996)
PubMed id: 8688419  
 
 
Solution structure of an engineered insulin monomer at neutral pH.
H.B.Olsen, S.Ludvigsen, N.C.Kaarsholm.
 
  ABSTRACT  
 
Insulin circulates in the bloodstream and binds to its specific cell-surface receptor as a 5808 Da monomeric species. However, studies of the monomer structure and dynamics in solution are severely limited by insulin self-association into dimers and higher oligomers. In the present work we use site-directed mutagenesis of the dimer- and hexamer-forming surfaces to yield the first insulin species amenable for structure determination at neutral pH by nuclear magnetic resonance (NMR) spectroscopy. The preferred insulin mutant, i.e., (B1, B10, B16, B27) Glu, des-B30 insulin retains 47% biological potency and remains monomeric at millimolar concentrations in aqueous solution at pH 6.5-7.5 as judged by NMR and near-UV circular dichroism (CD) spectroscopy. From a series of 2D 1H-NMR spectra collected at pH 6.5 and 34 degrees C, the majority of the resonances are assigned to specific residues in the sequence, and nuclear Overhauser enhancement (NOE) cross-peaks are identified. NOE-derived distance restraints in conjunction with torsion restraints based on measured coupling constants, 3JHNH alpha, are used for structure calculations using the hybrid method of distance geometry and simulated annealing. The calculated structures show that the major part of the insulin mutant is structurally well defined with an average root mean square (rms) deviation between the 25 calculated structures and the mean coordinates of 0.66 A for backbone atoms (A2-A19 and B4-B26) and 1.31 A for all backbone atoms. The A-chain consists of two antiparallel helices, A2-A7 and A12-A19, connected by a loop. The B-chain contains a loop region (B1-B8), an alpha-helix (B9-B19), and a type I turn (B20-B23) and terminates as an extended strand (B24-B29). The B1-B4 and B27-B29 regions are disordered in solution. The structure is generally similar to crystal structures and resembles a crystalline T-state more than an R-state in the sense that the B-chain helix is confined to residues B9-B19.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
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.  
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
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
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.  
18004974 Z.Y.Guo, Z.S.Qiao, and Y.M.Feng (2008).
The in vitro oxidative folding of the insulin superfamily.
  Antioxid Redox Signal, 10, 127-140.  
18085545 G.Grasso, E.Rizzarelli, and G.Spoto (2007).
AP/MALDI-MS complete characterization of the proteolytic fragments produced by the interaction of insulin degrading enzyme with bovine insulin.
  J Mass Spectrom, 42, 1590-1598.  
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
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
16762918 Q.X.Hua, S.Nakagawa, S.Q.Hu, W.Jia, S.Wang, and M.A.Weiss (2006).
Toward the active conformation of insulin: stereospecific modulation of a structural switch in the B chain.
  J Biol Chem, 281, 24900-24909.
PDB codes: 2hh4 2hho
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.  
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.  
15502368 H.Oneda, and K.Inouye (2004).
Detection of antigen-antibody reaction using a fluorescent probe and its application to homogeneous competitive-type immunoassay for insulin.
  Biosci Biotechnol Biochem, 68, 2190-2192.  
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
12723938 H.J.Choi, H.D.Je, J.H.Jeong, Y.S.Min, T.S.Choi, J.H.Park, C.Y.Shin, and U.D.Sohn (2003).
The role of ascorbic acid on the redox status and the concentration of malondialdehyde in streptozotocin-induced diabetic rats.
  Arch Pharm Res, 26, 237-243.  
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
12624089 Z.S.Qiao, C.Y.Min, Q.X.Hua, M.A.Weiss, and Y.M.Feng (2003).
In vitro refolding of human proinsulin. Kinetic intermediates, putative disulfide-forming pathway folding initiation site, and potential role of C-peptide in folding process.
  J Biol Chem, 278, 17800-17809.  
12081626 F.Y.Dupradeau, T.Richard, G.Le Flem, H.Oulyadi, Y.Prigent, and J.P.Monti (2002).
A new B-chain mutant of insulin: comparison with the insulin crystal structure and role of sulfonate groups in the B-chain structure.
  J Pept Res, 60, 56-64.
PDB code: 1ho0
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
11814349 Z.Y.Guo, L.Shen, and Y.M.Feng (2002).
The different folding behavior of insulin and insulin-like growth factor 1 is mainly controlled by their B-chain/domain.
  Biochemistry, 41, 1556-1567.  
12186542 Z.Y.Guo, L.Shen, and Y.M.Feng (2002).
The different energetic state of the intra A-chain/domain disulfide of insulin and insulin-like growth factor 1 is mainly controlled by their B-chain/domain.
  Biochemistry, 41, 10585-10592.  
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
10889022 G.M.Lipkind, and H.A.Fozzard (2000).
KcsA crystal structure as framework for a molecular model of the Na(+) channel pore.
  Biochemistry, 39, 8161-8170.  
11009601 H.B.Olsen, and N.C.Kaarsholm (2000).
Structural effects of protein lipidation as revealed by LysB29-myristoyl, des(B30) insulin.
  Biochemistry, 39, 11893-11900.  
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.  
  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
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
9631292 G.Dodson, and D.Steiner (1998).
The role of assembly in insulin's biosynthesis.
  Curr Opin Struct Biol, 8, 189-194.  
9733769 M.Jansson, G.Andersson, M.Uhlén, B.Nilsson, and J.Kördel (1998).
The insulin-like growth factor (IGF)binding protein 1 binding epitope on IGF-I probed by heteronuclear NMR spectroscopy and mutational analysis.
  J Biol Chem, 273, 24701-24707.  
9566117 C.McInnes, and B.D.Sykes (1997).
Growth factor receptors: structure, mechanism, and drug discovery.
  Biopolymers, 43, 339-366.  
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.