The minimal active domain (GEF domain) of the mouse Ras exchange factor CDC25(Mm) was purified to homogeneity from recombinant Escherichia coli culture. The 256 amino acids polypeptide shows high activity in vitro and forms a stable complex with H-ras p21 in absence of guanine nucleotides. Circular dichroism (CD) spectra in the far UV region indicate that this domain is highly structured with a high content of alpha-helix (42%). Near UV CD spectra evidenced good signal due to phenylalanine and tyrosine while a poor contribution was elicited by the three tryptophan residues contained in this domain. The tryptophan fluorescence signal was scarcely affected by denaturation of the protein or by formation of the binary complex with H-ras p21, suggesting that the Trp residues, which are well conserved in the GEF domain of several Ras-exchange factors, were exposed to the surface of the protein and they are not most probably directly involved in the interaction with Ras proteins

Coccetti, P., Monzani, E., Alberghina, L., Casella, L., & Martegani, E. (1998). Analysis of the secondary structure of the catalytic domain of mouse Ras exchange factor CDC25 Mm. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1383(2), 292-300 [10.1016/S0167-4838(97)00212-4].

Analysis of the secondary structure of the catalytic domain of mouse Ras exchange factor CDC25 Mm

COCCETTI, PAOLA;ALBERGHINA, LILIA;MARTEGANI, ENZO
1998

Abstract

The minimal active domain (GEF domain) of the mouse Ras exchange factor CDC25(Mm) was purified to homogeneity from recombinant Escherichia coli culture. The 256 amino acids polypeptide shows high activity in vitro and forms a stable complex with H-ras p21 in absence of guanine nucleotides. Circular dichroism (CD) spectra in the far UV region indicate that this domain is highly structured with a high content of alpha-helix (42%). Near UV CD spectra evidenced good signal due to phenylalanine and tyrosine while a poor contribution was elicited by the three tryptophan residues contained in this domain. The tryptophan fluorescence signal was scarcely affected by denaturation of the protein or by formation of the binary complex with H-ras p21, suggesting that the Trp residues, which are well conserved in the GEF domain of several Ras-exchange factors, were exposed to the surface of the protein and they are not most probably directly involved in the interaction with Ras proteins
No
Articolo in rivista - Articolo scientifico
Scientifica
Ras, catalytic domain
English
292
300
9
Coccetti, P., Monzani, E., Alberghina, L., Casella, L., & Martegani, E. (1998). Analysis of the secondary structure of the catalytic domain of mouse Ras exchange factor CDC25 Mm. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1383(2), 292-300 [10.1016/S0167-4838(97)00212-4].
Coccetti, P; Monzani, E; Alberghina, L; Casella, L; Martegani, E
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/10281/3519
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