Abstract
Integrin ?v?6 is a cell surface arginine-glycine-aspartic acid (RGD)-specific heterodimeric glycoprotein that is only expressed on epithelia during processes of tissue remodelling, including cancer. The specificity and molecular nature of interactions toward this integrin are poorly understood and new insights into such processes are important to cell biologists and pharmaceutical drug discovery. This study demonstrates the application of quantitative two-dimensional saturation transfer (Q2DSTD) NMR to obtain precise details of peptide interactions with integrin ?v?6 and their correlation to specificity for the integrin. This approach highlights subtle but significant differences in ligand contact by three related 21-mer peptides: FMDV2, an ?v?6 specific peptide and DBD1 and LAP2T1 peptides that bind many ?v integrins in addition to ?v?6. FMDV2 and DBD1 differ only by the cyclisation of DBD1; a process that removes ?v?6 specificity. Q2DSTD NMR demonstrates these peptides experience significantly different interactions with the integrin; FMDV contacts primarily through four residues: 6Leu, 10Leu, 12Val and 13Leu, whereas DBD1 and LAP2T1 have more widespread contacts across their sequences. Q2DSTD NMR combined two-dimensional STD with quantitation by considering the relaxation of the ligand (CRL) to provide precise ligand contact information. This study also examines the role of CRL in the Q2DSTD process and how quantitation modifies STD data and unravels epitope-mapping variability to provide precise results that differentiate interactions at the atomic level for each peptide.
[NMR paper] Epitope mapping by solution NMR spectroscopy.
Epitope mapping by solution NMR spectroscopy.
Epitope mapping by solution NMR spectroscopy.
J Mol Recognit. 2015 Feb 27;
Authors: Bardelli M, Livoti E, Simonelli L, Pedotti M, Moraes A, Valente AP, Varani L
Abstract
Antibodies play an ever more prominent role in basic research as well as in the biotechnology and pharmaceutical sectors. Characterizing their epitopes, that is, the region that they recognize on their target molecule, is useful for purposes ranging from molecular biology research to vaccine design and intellectual...
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03-03-2015 08:34 PM
[NMR paper] Determination of the binding epitope of RGD-peptidomimetics to ?v?3 and ?(IIb)?3 integrin-rich intact cells by NMR and computational studies.
Determination of the binding epitope of RGD-peptidomimetics to ?v?3 and ?(IIb)?3 integrin-rich intact cells by NMR and computational studies.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.rsc.org-images-entities-char_z_RSClogo.gif Related Articles Determination of the binding epitope of RGD-peptidomimetics to ?v?3 and ?(IIb)?3 integrin-rich intact cells by NMR and computational studies.
Org Biomol Chem. 2013 Jun 21;11(23):3886-93
Authors: Guzzetti I, Civera M, Vasile F, Araldi EM, Belvisi L, Gennari C, Potenza D, Fanelli R,...
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03-19-2014 10:43 PM
[NMR paper] Precise epitope mapping of malaria parasite inhibitory antibodies by TROSY NMR cross-
Precise epitope mapping of malaria parasite inhibitory antibodies by TROSY NMR cross-saturation.
Related Articles Precise epitope mapping of malaria parasite inhibitory antibodies by TROSY NMR cross-saturation.
Biochemistry. 2005 Jan 18;44(2):518-23
Authors: Morgan WD, Frenkiel TA, Lock MJ, Grainger M, Holder AA
We have applied NMR cross-saturation with TROSY detection to the problem of precisely mapping conformational epitopes on complete protein antigen molecules. We have investigated complexes of the Fab fragments of two antibodies that...
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11-24-2010 11:14 PM
[NMR paper] Epitope mapping of the phosphorylation motif of the HIV-1 protein Vpu bound to the se
Epitope mapping of the phosphorylation motif of the HIV-1 protein Vpu bound to the selective monoclonal antibody using TRNOESY and STD NMR spectroscopy.
Related Articles Epitope mapping of the phosphorylation motif of the HIV-1 protein Vpu bound to the selective monoclonal antibody using TRNOESY and STD NMR spectroscopy.
Biochemistry. 2004 Nov 23;43(46):14555-65
Authors: Gharbi-Benarous J, Bertho G, Evrard-Todeschi N, Coadou G, Megy S, Delaunay T, Benarous R, Girault JP
The conformational preferences of a 22-amino acid peptide...
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11-24-2010 10:03 PM
[NMR paper] Epitope mapping and competitive binding of HSA drug site II ligands by NMR diffusion
Epitope mapping and competitive binding of HSA drug site II ligands by NMR diffusion measurements.
Related Articles Epitope mapping and competitive binding of HSA drug site II ligands by NMR diffusion measurements.
J Am Chem Soc. 2004 Nov 3;126(43):14258-66
Authors: Lucas LH, Price KE, Larive CK
It is important to characterize drug-albumin binding during drug discovery and lead optimization as strong binding may reduce bioavailability and/or increase the drug's in vivo half-life. Despite knowing about the location of human serum albumin (HSA)...
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11-24-2010 10:03 PM
[NMR paper] Epitope mapping of sialyl Lewis(x) bound to E-selectin using saturation transfer diff
Epitope mapping of sialyl Lewis(x) bound to E-selectin using saturation transfer difference NMR experiments.
Related Articles Epitope mapping of sialyl Lewis(x) bound to E-selectin using saturation transfer difference NMR experiments.
Glycobiology. 2003 Jun;13(6):435-43
Authors: Rinnbauer M, Ernst B, Wagner B, Magnani J, Benie AJ, Peters T
A complex between sialyl Lewisx (alpha-D-Neu5Ac-- beta-D-Gal---beta-D-GlcNAc-O-8 COOMe) and E-selectin was studied using saturation transfer difference (STD) nuclear magnetic resonance (NMR) experiments....
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11-24-2010 09:01 PM
[NMR paper] Epitope mapping of ligand-receptor interactions by diffusion NMR.
Epitope mapping of ligand-receptor interactions by diffusion NMR.
Related Articles Epitope mapping of ligand-receptor interactions by diffusion NMR.
J Am Chem Soc. 2002 Aug 28;124(34):9984-5
Authors: Yan J, Kline AD, Mo H, Zartler ER, Shapiro MJ
A novel method based on diffusion NMR for the epitope mapping of ligand binding is presented. The intermolecular NOE builds up during a long diffusion period and creates a deviation from the linearity. The ligand proton nearest the protein generates the strongest NOE from protein during the diffusion...
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11-24-2010 08:58 PM
[NMR paper] Group epitope mapping by saturation transfer difference NMR to identify segments of a
Group epitope mapping by saturation transfer difference NMR to identify segments of a ligand in direct contact with a protein receptor.
Related Articles Group epitope mapping by saturation transfer difference NMR to identify segments of a ligand in direct contact with a protein receptor.
J Am Chem Soc. 2001 Jun 27;123(25):6108-17
Authors: Mayer M, Meyer B
A protocol based on saturation transfer difference (STD) NMR spectra was developed to characterize the binding interactions at an atom level, termed group epitope mapping (GEM). As an example...
Q2DSTD NMR deciphers epitope-mapping variability for peptide recognition of integrin ?v?6.
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