Conformational flexibility in a small globular hormone: X‐ray analysis of avian pancreatic polypeptide at 0.98‐Å resolution

Glover, I. D.; Haneef, I.; Pitts, John; Wood, Steve; Moss, David S.; Tickle, Ian J.; Blundell, Tom L.

doi:10.1002/bip.360220138

Cited by 221 publications

(88 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Each of these models confirms a hairpin structure of neuropeptide Y for its binding conformations to Y, and Y, receptors. The model described in this paper was built by homology to the X-ray structure of avian pancreatic peptide (Glover et al, 1983) obtained from Protein Data Bank entry 1PPT. The hairpin structure is represented by a type I1 proline helix adopted by the residues 1-8, a ,&turn structure made of amino acids 9 -14 and an amphiphilic a-helical segment comprising the residues 15-32.…”

Section: Molecular Modelling Of Neuropeptide Ymentioning

confidence: 99%

Complete L‐Alanine Scan of Neuropeptide Y Reveals Ligands Binding to Y₁ and Y₂ Receptors with Distinguished Conformations

Beck‐Sickinger

Weland

Wittneben

et al. 1994

European Journal of Biochemistry

178

223

View full text Add to dashboard Cite

The synthesis of more than fifty 36-residue oligopeptide analogs of neuropeptide Y (NPY) and their affinity to human Y, and Y, receptors is described. Each amino acid of the natural sequence was replaced by L-alanine, the four alanine residues at position 12, 14, 18 and 23 were replaced by glycine. Additional residues .were exchanged to closely related ones in order to characterize the prerequisites for binding. A combination of automated single and multiple peptide synthesis using fluoren-9-ylmethoxycarbonyl/tert-butoxy strategy was applied. The purified peptides were characterized by electrospray mass spectrometry, analytical HPLC and amino acid analysis. Binding was investigated by displacement of '251-labelled neuropeptide Y from human neuroblastoma cell lines Whereas Pro2 and the integrity of the neuropeptide Y loop is important for the binding to the Y, receptor, exchanges within the C-terminal helix affect the affinity to the Y, receptor. The Cterminal pentapeptide amide is important for both receptors and probably represents the binding site. However, Arg33 and Arg35 may not be exchanged by L-alanine in the Y, system, whereas Arg35 and Tyr36 are the most susceptible residues in the Y, system. In order to distinguish between conformational effects and direct hormone/receptor interaction via the side chains of neuropeptide Y, circular dichroic studies of the alanine-containing peptides were performed and structure affinity relationships are discussed.Comparing the affinities of the neuropeptide Y analogs to Y, and Y, receptors significant differences were found for the two binding sites, which suggests a different active conformation of neuropeptide Y at the two subtypes of receptors. Using molecular dynamics calculations, two distinct conformations were identified which are in good agreement with the data obtained by structure/ affinity investigations. SK-N-MC and SMS-KAN.Neuropeptide Y (NPY) is a 36-amino-acid amide which exhibits a high similarity to the pancreatic polypeptide and peptide YY. It was isolated from pig brain and sequenced (Tatemoto) in 1982.Information about the secondary structure of pancreatic polypetptide hormones has been obtained through the work of Blundell and co-workers who performed X-ray analyses on avian (turkey) pancreatic polypeptide Wood, 1982., Glover et al., 1983). Residues 1-8 of the crystalline avian pancreatic polypeptide form a type I1 proline helix followed by a loop (residues 9-14) and an a-helical segment (residues 15-32). The four C-terminal amino acids adopt a relatively flexible loop-like conformation. Hydrophobic interactions of the proline helix and the amphiphilic ahelix lead to the hairpin structure. Comparative circular dichroic measurements of different pancreatic polypeptides and Correspondence to A. G.

show abstract

Section: Molecular Modelling Of Neuropeptide Ymentioning

confidence: 99%

Complete L‐Alanine Scan of Neuropeptide Y Reveals Ligands Binding to Y₁ and Y₂ Receptors with Distinguished Conformations

Beck‐Sickinger

Weland

Wittneben

et al. 1994

European Journal of Biochemistry

178

223

View full text Add to dashboard Cite

show abstract

“…Its crystal structure was determined by Blundell et al (1981) at 1.4 A resolution and then by Glover et al (1983) at a resolution of 0.98 A. The structure consists of a polyproline-like or collagen-like helix running from residues 1 to 8, packed against the hydrophobic face of an a-helix that extends from residues 13 to 3 1.…”

Section: Test Proteinmentioning

confidence: 99%

Prediction of protein conformation on the basis of a search for compact structures: Test on avian pancreatic polypeptide

et al. 1993

View full text Add to dashboard Cite

Based on the concept that hydrophobic interactions cause a polypeptide chain to adopt a compact structure, a method is proposed to predict the structure of a protein. The procedure is carried out in four stages: (1) use of a virtual-bond united-residue approximation with the side chains represented by spheres to search conformational space extensively using specially designed interactions to lead to a collapsed structure, (2) conversion of the lowest-energy virtual-bond united-residue chain to one with a real polypeptide backbone, with optimization of the hydrogen-bond network among the backbone groups, (3) perturbation of the latter structure by the electrostatically driven Monte Carlo (EDMC) procedure, and (4) conversion of the spherical representation of the side chains to real groups and perturbation of the whole molecule by the EDMC procedure using the empirical conformational energy program for peptides (ECEPP/2) energy function plus hydration. Application of this procedure to the 36-residue avian pancreatic polypeptide led to a structure that resembled the one determined by X-ray crystallography; it had an a-helix starting at residue 13, with the N-terminal portion of the chain in an extended conformation packed against the a-helix. Similar structures with slightly higher energies, but looser packing, were also obtained.Keywords: compact conformations; conversion from a united-residue representation to an all-atom chain; hydrophobic-residue packing; Monte Carlo methods; multiple-minima problem; potential of mean force; protein folding; united-residue representation of a polypeptide chain In our continuing effort to surmount the multiple-minima problem (Scheraga, 1989;Kostrowicki & Scheraga, 1992;Olszewski et al., 1992) in computing the structure of a protein, we have developed a procedure that takes advantage of the fact that the protein core tends to consist of tightly packed nonpolar residues, with the polar ones located on the surface (Kauzmann, 1959;Rackovsky & Scheraga, 1977;Richards, 1977;Wertz & Scheraga, 1978;Chan & Dill, 1990;. Such an example of selforganization is reminiscent of early work by Onsager (1949) on the packing of tobacco mosaic virus particles and by Flory (1956) on the packing of rodlike polymers (including a-helices). These workers showed that, as the solution concentration increases, the system separates into two phases, an organized ordered one and a more dilute isotropic one. The self-organization of the moreconcentrated ordered phase arises solely from entropic effects; i.e., in a concentrated solution it is easier to pack rods in an ordered anisotropic array than in a disordered isotropic one. Of course, while entropy alone can account for this phenomenon, possible attractive interactions between the ordered rods can also contribute to this selforganization. We thus consider the possibility that, if the available conformational space of a polypeptide is confined, organized structure will be promoted.

show abstract

“…Neuropeptide Y (NPY) is a 36-peptide amide which exhibits a high homology with the pancreatic polypeptide (PP) and peptide YY (PYY) in both sequence and 3D structure [1,2]. It was isolated from pig brain and sequenced in 1982 [3].…”

Section: Introductionmentioning

confidence: 99%

Modified, cyclic dodecapeptide analog of neuropeptide Y is the smallest full agonist at the human Y₂ receptor

et al. 1996

View full text Add to dashboard Cite

In order to stabilize the C-terminal dodecapeptide of neuropeptide Y (NPY) we replaced Leu 2s and Thr 32 by Lys and Glu, respectively, and subsequently linked these residues by lactamization. This peptide analog of NPY shows a more than 100-fold increase in affinity compared to the C-terminal linear dodecapeptide in receptor binding studies performed at human neuroblastoma cells SMS-KAN, which exclusively express the Y2 receptor subtype.2+ Signal transduction was investigated" by measuring Ca current inhibition in human SH-SY5Y cells and cyclic [Lys2S-Glu3Z] NPY Ac-25-36 and NPY were shown to be equipotent in this assay. Thus, this molecule is the smallest Y2 receptor selective full agonist of NPY. Using 2D-NMR experiments and molecular modelling techniques, the structures of the linear and cyclic peptides have been investigated and significant differences have been found, which may explain the improvement in biological activity. Thus, a model of the bioactive conformation of NPY at the human Yz receptor is suggested.

show abstract

Conformational flexibility in a small globular hormone: X‐ray analysis of avian pancreatic polypeptide at 0.98‐Å resolution

Cited by 221 publications

References 14 publications

Complete L‐Alanine Scan of Neuropeptide Y Reveals Ligands Binding to Y₁ and Y₂ Receptors with Distinguished Conformations

Complete L‐Alanine Scan of Neuropeptide Y Reveals Ligands Binding to Y₁ and Y₂ Receptors with Distinguished Conformations

Prediction of protein conformation on the basis of a search for compact structures: Test on avian pancreatic polypeptide

Modified, cyclic dodecapeptide analog of neuropeptide Y is the smallest full agonist at the human Y₂ receptor

Contact Info

Product

Resources

About

Conformational flexibility in a small globular hormone: X‐ray analysis of avian pancreatic polypeptide at 0.98‐Å resolution

Cited by 221 publications

References 14 publications

Complete L‐Alanine Scan of Neuropeptide Y Reveals Ligands Binding to Y1 and Y2 Receptors with Distinguished Conformations

Complete L‐Alanine Scan of Neuropeptide Y Reveals Ligands Binding to Y1 and Y2 Receptors with Distinguished Conformations

Prediction of protein conformation on the basis of a search for compact structures: Test on avian pancreatic polypeptide

Modified, cyclic dodecapeptide analog of neuropeptide Y is the smallest full agonist at the human Y2 receptor

Contact Info

Product

Resources

About

Complete L‐Alanine Scan of Neuropeptide Y Reveals Ligands Binding to Y₁ and Y₂ Receptors with Distinguished Conformations

Complete L‐Alanine Scan of Neuropeptide Y Reveals Ligands Binding to Y₁ and Y₂ Receptors with Distinguished Conformations

Modified, cyclic dodecapeptide analog of neuropeptide Y is the smallest full agonist at the human Y₂ receptor