Keywords: Cd Spectroscopy, Domain Refolding, Enzyme Digestion, Peptide Synthesis, Receptor Binding, Growth Factor, Protein Cripto, Unclassified Drug, Article, Circular Dichroism, Conformational Transition, Molecular Recognition, Mouse, Nonhuman, Priority Journal, Protein Domain, Protein Synthesis, Activin Receptors, Type I, Amino Acid Sequence, Animals, Binding Sites, Chromatography, Liquid, Epidermal Growth Factor, Kinetics, Mass Spectrometry, Membrane Glycoproteins, Models, Molecular Sequence Data, Mutation, Neoplasm Proteins, Protein Binding, Protein Folding, Protein Structure, Tertiary, Recombinant Proteins, Sequence Homology,
Affiliations: *** IBB - CNR ***
Istituto di Biostrutture e Bioimmagini del CNR, Sezione Biostrutture, Napoli, Italy
Istituto di Genetica e Biofisica Adriano B. Traverso del CNR, Napoli, Italy
Facoltà di Scienze MMFFNN, Seconda Università di Napoli, Caserta, Italy
Facolt di Scienze MMFFNN, Seconda Universit di Napoli, Caserta, Italy
References: Not available.
Chemical synthesis of mouse Cripto CFC variants
We report for the first time the chemical synthesis of refolded CFC domain of mouse Cripto (mCFC) and of two variants bearing mutations on residues W107 and H104 involved in Alk4 binding. The domains undergo spontaneous and quantitative refolding in about 4 h, yet with very different kinetics. Disulfide linkages have been assessed by enzyme digestion and mass spectrometry analysis of resulting fragments, and the first experimental studies on structural organization have been conducted by circular dichroism spectroscopy under different pH conditions. Upon refolding, the domains considerably change their conformations, although they do not assume canonical structures, and become highly resistant to enzyme degradation. A comparative study of receptor binding shows that the CFC domain can bind Alk4 and confirms the importance of W107 and H104 for receptor recognition.
Aloj L, Aurilio M, Rinaldi V, D'Ambrosio L, Tesauro D, Peitl PK, Maina T, Mansi R, Von Guggenberg E, Joosten L, Sosabowski JK, Breeman WA, De Blois E, Koelewijn S, Melis M, Waser B, Beetschen K, Reubi JC, De Jong M * The EEE project(453 views) Proc Int Cosm Ray Conf Icrc Universidad Nacional Autonoma De Mexico, 2007; 5(HEPART2): 977-980. Impact Factor:0 ViewExport to BibTeXExport to EndNote