Related ArticlesStructural investigations of a human calcitonin-derived carrier peptide in a membrane environment by solid-state NMR.
Biochemistry. 2004 Oct 5;43(39):12459-68
Authors: Wagner K, Beck-Sickinger AG, Huster D
Previous studies have shown that human calcitonin (hCT) and its C-terminal fragment hCT(9-32) translocate in nasal epithelium. Moreover, hCT(9-32) was used as a carrier to internalize efficiently the green fluorescent protein, drugs, and plasmid DNA. To understand the mechanism of the membrane crossing process, we determined structural parameters of the carrier peptide hCT(9-32) in a membrane environment using solid-state NMR. For that purpose, we synthesized a multiply labeled hCT(9-32) peptide comprising four positions with fully (15)N- and (13)C-labeled amino acids. Multilamellar vesicle samples containing varying mixing ratios of hCT(9-32) and phospholipids found in the plasma membrane of nasal epithelium were prepared. The typical axially symmetric powder patterns of (31)P NMR spectra confirmed the presence of lamellar bilayers in our samples. The chemical shift anisotropy of the (31)P NMR spectra of the samples in the presence of hCT(9-32) is slightly reduced, revealing weak interaction of the peptide with the lipid headgroups. The peptide does not penetrate the lipid membrane as indicated by very similar (2)H NMR order parameters of the phospholipid fatty acid chains in the absence and presence of the carrier peptide. This membrane topology was confirmed by measurements of paramagnetic enhancement of relaxation rates. The conformation of hCT(9-32) was investigated by cross polarization magic angle spinning NMR methods. All peptide signals were resolved and fully assigned in two-dimensional proton-driven (13)C spin diffusion experiments. The isotropic chemical shifts of (13)CO, (13)Calpha, and (13)Cbeta provide information about the secondary structure of the carrier peptide. The conformation of hCT(9-32) was further corroborated by quantitative phi torsion angle measurements. Two monomeric structural models are consistent with the data: (i) a linear backbone conformation of hCT(9-32) and (ii) an antiparallel beta-sheet structure. These structures are maintained over a wide range of peptide:lipid mixing ratios. No direct indications for fibril formation of hCT(9-32) were found. Dipolar coupling measurements indicate rather high amplitudes of motion of the peptide.
Expression and purification of (15)N- and (13)C-isotope labeled 40-residue human Alzheimer's ?-amyloid peptide for NMR-based structural analysis.
Expression and purification of (15)N- and (13)C-isotope labeled 40-residue human Alzheimer's ?-amyloid peptide for NMR-based structural analysis.
Expression and purification of (15)N- and (13)C-isotope labeled 40-residue human Alzheimer's ?-amyloid peptide for NMR-based structural analysis.
Protein Expr Purif. 2011 May 27;
Authors: Long F, Cho W, Ishii Y
Amyloid fibrils of Alzheimer's ?-amyloid peptide (A?) are a primary component of amyloid plaques, a hallmark of Alzheimer's disease (AD). Enormous attention has been given to the structural...
nmrlearner
Journal club
0
06-07-2011 11:05 AM
[NMR paper] NMR investigations of the role of the sugar moiety in glycosylated recombinant human
NMR investigations of the role of the sugar moiety in glycosylated recombinant human granulocyte-colony-stimulating factor.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif Related Articles NMR investigations of the role of the sugar moiety in glycosylated recombinant human granulocyte-colony-stimulating factor.
Eur J Biochem. 1997 Jul 1;247(1):386-95
Authors: Gervais V, Zerial A, Oschkinat H
Human granulocyte-colony-stimulating factor (G-CSF) is a...
nmrlearner
Journal club
0
08-22-2010 03:31 PM
[NMR paper] NMR investigations of the role of the sugar moiety in glycosylated recombinant human
NMR investigations of the role of the sugar moiety in glycosylated recombinant human granulocyte-colony-stimulating factor.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif Related Articles NMR investigations of the role of the sugar moiety in glycosylated recombinant human granulocyte-colony-stimulating factor.
Eur J Biochem. 1997 Jul 1;247(1):386-95
Authors: Gervais V, Zerial A, Oschkinat H
Human granulocyte-colony-stimulating factor (G-CSF) is a...
nmrlearner
Journal club
0
08-22-2010 03:03 PM
[NMR paper] NMR investigations of the structural properties of the nodulation protein, NodF, from
NMR investigations of the structural properties of the nodulation protein, NodF, from Rhizobium leguminosarum and its homology with Escherichia coli acyl carrier protein.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles NMR investigations of the structural properties of the nodulation protein, NodF, from Rhizobium leguminosarum and its homology with Escherichia coli acyl carrier protein.
FEBS Lett. 1996 Jun 10;388(1):66-72
Authors: Ghose R, Geiger O, Prestegard JH
...
nmrlearner
Journal club
0
08-22-2010 02:27 PM
[NMR paper] What function for human lithostathine?: structural investigations by three-dimensiona
What function for human lithostathine?: structural investigations by three-dimensional structure modeling and high-resolution NMR spectroscopy.
Related Articles What function for human lithostathine?: structural investigations by three-dimensional structure modeling and high-resolution NMR spectroscopy.
Protein Eng. 1996 Nov;9(11):949-57
Authors: Patard L, Stoven V, Gharib B, Bontems F, Lallemand JY, De Reggi M
Human lithostathine is a 144-residue protein, expressed in various organs and pathologies. Several biological functions have been...
[NMR paper] NMR-derived model for a peptide-antibody complex.
NMR-derived model for a peptide-antibody complex.
Related Articles NMR-derived model for a peptide-antibody complex.
Biochemistry. 1990 Oct 30;29(43):10032-41
Authors: Zilber B, Scherf T, Levitt M, Anglister J
The TE34 monoclonal antibody against cholera toxin peptide 3 (CTP3; VEVPGSQHIDSQKKA) was sequenced and investigated by two-dimensional transferred NOE difference spectroscopy and molecular modeling. The VH sequence of TE34, which does not bind cholera toxin, shares remarkable homology to that of TE32 and TE33, which are both anti-CTP3...