Biochemical dissection of Anosmin-1 interaction with FGFR1 and components of the extracellular matrix.
Anosmin-1, defective in Kallmann's syndrome, participates in the adhesion, migration and differentiation of different cell types in the CNS. Although not fully understood, the mechanisms of action of Anosmin-1 involve the interaction with different proteins, being the interaction with fibroblast growth factor receptor 1 (FGFR1) and the modulation of its signalling the best studied to date. Using glutathione-S-transferase pull-down assays we demonstrate that the FnIII.3 (Fibronectin-like type III) domain and the combination whey acidic protein-FnIII.1, but not each of them individually, interact with FGFR1. The interaction of the whey acidic protein-FnIII.1 domains is substantially reduced when the cysteine-rich region is present, suggesting a likely regulatory role for this domain. The introduction in FnIII.3 of any of the two missense mutations found in Kallmann's syndrome patients, E514K and F517L, abolished the interaction with FGFR1, what suggests an important role for these residues in the interaction. Interestingly, the chemoattraction of Anosmin-1 on rat neuronal precursors (NPs) via FGFR1 is retained by the N-terminal region of Anosmin-1 but not by FnIII.3 alone, and is lost in proteins carrying either one of the missense mutations, probably because of a highly reduced binding capacity to FGFR1. We also describe homophilic interaction Anosmin-1/Anosmin-1 via the FnIII repeats 1 and 4, and the interaction of FnIII.1 and FnIII.3 with Fibronectin and of FnIII.3 with Laminin.
Murcia-Belmonte
Verónica
V
Grupo de Neurobiología del Desarrollo-GNDe, Hospital Nacional de Parapléjicos, Toledo, Spain.
Esteban
Pedro F
PF
García-González
Diego
D
De Castro
Fernando
F
D000818:Animals; D000831:Animals, Newborn; D016466:CHO Cells; D058565:Cerebral Ventriculitis; D002633:Chemotaxis / Q000235:genetics / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005109:Extracellular Matrix / Q000737:chemistry* / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D007797:Laminin; D009154:Mutation / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D058953:Neural Stem Cells / Q000187:drug effects / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D011758:Pyrroles / Q000494:pharmacology; D051381:Rats; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods
C076266:ANOS1 protein, human; D017077:Culture Media, Conditioned; D004791:Enzyme Inhibitors; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D007797:Laminin; D009419:Nerve Tissue Proteins; D011758:Pyrroles; C105686:SU 5402; D051496:Receptor, Fibroblast Growth Factor, Type 1; D003545:Cysteine