[NMR paper] Combined ligand-observe (19)F and protein-observe (15)N,(1)H-HSQC NMR suggests phenylalanine as the key ?-somatostatin residue recognized by human protein disulfide isomerase.
Combined ligand-observe (19)F and protein-observe (15)N,(1)H-HSQC NMR suggests phenylalanine as the key ?-somatostatin residue recognized by human protein disulfide isomerase.
Combined ligand-observe (19)F and protein-observe (15)N,(1)H-HSQC NMR suggests phenylalanine as the key ?-somatostatin residue recognized by human protein disulfide isomerase.
Abstract
Human protein disulphide isomerase (hPDI) is an endoplasmic reticulum (ER) based isomerase and folding chaperone. Molecular detail of ligand recognition and specificity of hPDI are poorly understood despite the importance of the hPDI for folding secreted proteins and its implication in diseases including cancer and lateral sclerosis. We report a detailed study of specificity, interaction and dissociation constants (Kd) of the peptide-ligand ?-somatostatin (AGSKNFFWKTFTSS) binding to hPDI using (19)F ligand-observe and (15)N,(1)H-HSQC protein-observe NMR methods. Phe residues in ?-somatostatin are hypothesised as important for recognition by hPDI therefore, step-wise peptide Phe-to-Ala changes were progressively introduced and shown to raise the Kd from 103 + 47 ?M until the point where binding was abolished when all Phe residues were modified to Ala. The largest step-changes in Kd involved the F11A peptide modification which implies the C-terminus of ?-somatostatin is a prime recognition region. Furthermore, this study also validated the combined use of (19)F ligand-observe and complimentary (15)N,(1)H-HSQC titrations to monitor interactions from the protein's perspective. (19)F ligand-observe NMR was ratified as mirroring (15)N protein-observe but highlighted the advantage that (19)F offers improved Kd precision due to higher spectrum resolution and greater chemical environment sensitivity.
[NMR paper] The PRE-Derived NMR Model of the 38.8kDa Tri-Domain IsdH Protein from Staphylococcus aureus Suggests that it Adaptively Recognizes Human Hemoglobin.
The PRE-Derived NMR Model of the 38.8kDa Tri-Domain IsdH Protein from Staphylococcus aureus Suggests that it Adaptively Recognizes Human Hemoglobin.
Related Articles The PRE-Derived NMR Model of the 38.8kDa Tri-Domain IsdH Protein from Staphylococcus aureus Suggests that it Adaptively Recognizes Human Hemoglobin.
J Mol Biol. 2015 Feb 13;
Authors: Sjodt M, Macdonald R, Spirig T, Chan AH, Dickson CF, Fabian M, Olson JS, Gell DA, Clubb RT
Abstract
Staphylococcus aureus is a medically important bacterial pathogen that during...
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The PRE-Derived NMR Model of the 38.8kDa Tri-Domain IsdH Protein from Staphylococcus aureus Suggests that it Adaptively Recognizes Human Hemoglobin
The PRE-Derived NMR Model of the 38.8kDa Tri-Domain IsdH Protein from Staphylococcus aureus Suggests that it Adaptively Recognizes Human Hemoglobin
Publication date: Available online 14 February 2015
Source:Journal of Molecular Biology</br>
Author(s): Megan Sjodt , Ramsay Macdonald , Thomas Spirig , Albert H. Chan , Claire F. Dickson , Marian Fabian , John S. Olson , David A. Gell , Robert T. Clubb</br>
Staphylococcus aureus is a medically important bacterial pathogen that during infections acquires iron from human hemoglobin (Hb). It uses two closely...
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02-14-2015 03:52 PM
[NMR paper] (19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b'x from human protein disulphide isomerase (hPDI).
(19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b'x from human protein disulphide isomerase (hPDI).
Related Articles (19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b'x from human protein disulphide isomerase (hPDI).
Org Biomol Chem. 2014 May 6;
Authors: Curtis-Marof R, Doko D, Rowe ML, Richards KL, Williamson RA, Howard MJ
Abstract
We report a protein-observe (19)F NMR-based ligand titration binding study of human PDI b'x with ?-somatostatin that also emphasises...
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05-08-2014 05:26 AM
Superconducting Magnets Used to Observe Proteins, Atom-by-Atom - Azom.com
Superconducting Magnets Used to Observe Proteins, Atom-by-Atom - Azom.com
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Superconducting Magnets Used to Observe Proteins, Atom-by-Atom
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â??This is a very specialized facility,â?? says Stanley Opella, professor of chemistry and biochemistry and director of the Center for NMR Spectroscopy and Imaging of Proteins, the organization that operates this building. The purpose of the facility is to ...
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10-27-2012 03:04 AM
[Question from NMRWiki Q&A forum] How to extract residue information in case of line broadening (intensity drop ) of HSQC titration of protein - protein interaction data ?
How to extract residue information in case of line broadening (intensity drop ) of HSQC titration of protein - protein interaction data ?
Dear Friends
we have done HSQC gradient titration with targeted proteins , most of cross peaks we observed intensity drop , 30 out of 95 peaks ( 70 percent intensity drop of each cross peak ) , other cross peaks also we observe intensity drop less than 70 percent , we did n"t observe any chemical shift change .( May be it is due to intermediate exchange regime of complex ) .ITC experiment showing 12uM binding constant .AUC experiment result is...
[NMR paper] Application of the 19F NMR technique to observe binding of the general anesthetic hal
Application of the 19F NMR technique to observe binding of the general anesthetic halothane to human serum albumin.
Related Articles Application of the 19F NMR technique to observe binding of the general anesthetic halothane to human serum albumin.
Anal Sci. 2004 Oct;20(10):1475-7
Authors: Shikii K, Sakurai S, Utsumi H, Seki H, Tashiro M
19F NMR techniques were employed to characterize the binding property of the widely used general anesthetic halothane with human serum albumin (HSA). It was found that 19F(1H) NOE and 2D 1H-19F HOESY...
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11-24-2010 10:01 PM
[NMR paper] NMR structures of three single-residue variants of the human prion protein.
NMR structures of three single-residue variants of the human prion protein.
Related Articles NMR structures of three single-residue variants of the human prion protein.
Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8340-5
Authors: Calzolai L, Lysek DA, Guntert P, von Schroetter C, Riek R, Zahn R, Wüthrich K
The NMR structures of three single-amino acid variants of the C-terminal domain of the human prion protein, hPrP(121-230), are presented. In hPrP(M166V) and hPrP(R220K) the substitution is with the corresponding residue in murine PrP, and in...
Combined ligand-observe (19)F and protein-observe (15)N,(1)H-HSQC NMR suggests phenylalanine as the key ?-somatostatin residue recognized by human protein disulfide isomerase.
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