title: A Combined NMR and Computational Approach to Determine the RGDechi-hCit-αv β3 Integrin Recognition Mode in Isolated Cell Membranes.
abstract: The critical role of integrins in tumor progression and metastasis has stimulated intense efforts to identify pharmacological agents that can modulate integrin function. In recent years, αv β3 and αv β5 integrin antagonists were demonstrated to be effective in blocking tumor progression. RGDechi-hCit, a chimeric peptide containing a cyclic RGD motif linked to an echistatin C-terminal fragment, is able to recognize selectively αv β3 integrin both in vitro and in vivo. High-resolution molecular details of the selective αv β3 recognition of the peptide are certainly required, nonetheless RGDechi-hCit internalization limited the use of classical in cell NMR experiments. To overcome such limitations, we used WM266 isolated cellular membranes to accomplish a detailed NMR interaction study that, combined with a computational analysis, provides significant structural insights into αv β3 molecular recognition by RGDechi-hCit. Remarkably, on the basis of the identified molecular determinants, we design a RGDechi-hCit mutant that is selective for αv β5 integrin.
authors.affiliation: Dipatimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, Seconda Università degli Studi Napoli, Via Vivaldi 46, 81100, Caserta (Italy).|Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, 80134 Naples (Italy).
authors.forename: Biancamaria
authors.initials: B
authors.lastname: Farina
authors.affiliation: Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, 80134 Naples (Italy).
authors.forename: Ivan
authors.initials: I
authors.lastname: de Paola
authors.affiliation: Dipatimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, Seconda Università degli Studi Napoli, Via Vivaldi 46, 81100, Caserta (Italy).
authors.forename: Luigi
authors.initials: L
authors.lastname: Russo
authors.affiliation: Dipartimento di Farmacia, Università di Napoli Federico II, Via Mezzocannone 16, 80134 Naples (Italy).
authors.forename: Domenica
authors.initials: D
authors.lastname: Capasso
authors.affiliation: Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, 80134 Naples (Italy).
authors.forename: Annamaria
authors.initials: A
authors.lastname: Liguoro
authors.affiliation: Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, 80134 Naples (Italy).
authors.forename: Annarita Del
authors.initials: AD
authors.lastname: Gatto
authors.affiliation: Istituto di Cristallografia, CNR, Via Amendola 122/O, 70126 Bari (Italy).
authors.forename: Michele
authors.initials: M
authors.lastname: Saviano
authors.affiliation: Dipatimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, Seconda Università degli Studi Napoli, Via Vivaldi 46, 81100, Caserta (Italy).
authors.forename: Paolo V
authors.initials: PV
authors.lastname: Pedone
authors.affiliation: Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, 80134 Naples (Italy).
authors.forename: Sonia
authors.initials: S
authors.lastname: Di Gaetano
authors.affiliation: Dipatimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, Seconda Università degli Studi Napoli, Via Vivaldi 46, 81100, Caserta (Italy).
authors.forename: Gaetano
authors.initials: G
authors.lastname: Malgieri
authors.affiliation: Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, 80134 Naples (Italy). lzaccaro@unina.it.
authors.forename: Laura
authors.initials: L
authors.lastname: Zaccaro
authors.affiliation: Dipatimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, Seconda Università degli Studi Napoli, Via Vivaldi 46, 81100, Caserta (Italy). roberto.fattorusso@unina2.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Fattorusso
mesh_terms: D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D039301:Computers, Molecular; D039302:Integrin alphaVbeta3 / Q000737:chemistry* / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D009842:Oligopeptides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D019030:Receptors, Vitronectin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D039302:Integrin alphaVbeta3; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D009842:Oligopeptides; D010455:Peptides; D019030:Receptors, Vitronectin; C064660:integrin alphaVbeta5; C057986:echistatin; C047981:arginyl-glycyl-aspartic acid
keywords: NMR spectroscopy; RGD ligand; cell membranes; integrin; molecular docking