Structural investigations on the Nodal-Cripto binding: a theoretical and experimental approach.
Nodal, a member of the transforming growth factor-β superfamily, is a potent embryonic morphogen also implicated in tumor progression. Up to date structural information on the interaction of Nodal with its molecular partners are unknown. To deepen our understanding about mechanisms underlying both embryonic development and Nodal/Cripto-dependent tumor progression, we present here a molecular model of activin receptor-like kinase 4/Cripto/Nodal complex built by homology modeling as well as docking tests aimed at identifying potential binding epitopes. Starting from this model, we have predicted a large interaction surface on Nodal, which encompasses residues 43-69 and includes the prehelix loop and the H3 helix. This hypothesis has been subsequently assessed by surface plasmon resonance binding assays between the full-length Cripto and synthetic peptides reproducing the selected Nodal regions. In addition, the binding affinity between the full-length Nodal and Cripto proteins has been evaluated for the first time.
Calvanese
Luisa
L
Department of Chemistry P. Corradini, University of Naples Federico II, Via Cintia 45, 80126 Naples, Italy.
Marasco
Daniela
D
Doti
Nunzianna
N
Saporito
Angela
A
D'Auria
Gabriella
G
Paolillo
Livio
L
Ruvo
Menotti
M
Falcigno
Lucia
L
D030201:Activin Receptors, Type I / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004815:Epidermal Growth Factor / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D058851:GPI-Linked Proteins; D006801:Humans; D066298:In Vitro Techniques; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D046912:Multiprotein Complexes / Q000737:chemistry; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055457:Nodal Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D020349:Surface Plasmon Resonance
D058851:GPI-Linked Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D046912:Multiprotein Complexes; C526007:NODAL protein, human; D009363:Neoplasm Proteins; D055457:Nodal Protein; D011994:Recombinant Proteins; C070636:TDGF1 protein, human; D004815:Epidermal Growth Factor; C498932:ACVR1B protein, human; D030201:Activin Receptors, Type I