The minimal active domain of the mouse CDC25Mm, a GDP/GTP exchange factor (GEF) active on H-ras protein, was determined by constructing several deletion mutants of the C-terminal domain of the protein. The functional activity of these fragments was analyzed for the ability to complement the yeast temperature sensitive mutation cdc25-1 and to catalyze the GDP/GTP exchange on Ras proteins in vitro. A C-terminal domain of 256 residues (CDC25 Mm1005-1260) was sufficient for full biological activity in vivo. Deletion of 27 C-terminal amino acids (CDC25 Mm1005-1233) abolished the complementing activity while deletion of 25 N-terminal residues (CDC25Mm1030-1260 corresponding to the most conserved domain) led to a complete loss of expression. The results in vivo were supported by experiments in vitro. Highly purified CDC25Mm1005-1260, expressed in E. coli using the pMAL system, enhanced the GDP release from both H-ras p21 and S. cerevisiae Ras2p and its activity was nearly as high as that of CDC25Mm974-1260. Comparison with the Cdc25p protein yielded further evidence that the minimal active domain of CDC25Mm is shorter than the yeast one. © 1995 Academic Press, Inc.
Coccetti, P., Mauri, I., Alberghina, L., Martegani, E., & Parmeggiani, A. (1995). The minimal active domain of the mouse Ras exchange factor CDC25Mm. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 206(1), 253-259 [10.1006/bbrc.1995.1035].
|Citazione:||Coccetti, P., Mauri, I., Alberghina, L., Martegani, E., & Parmeggiani, A. (1995). The minimal active domain of the mouse Ras exchange factor CDC25Mm. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 206(1), 253-259 [10.1006/bbrc.1995.1035].|
|Tipo:||Articolo in rivista - Articolo scientifico|
|Carattere della pubblicazione:||Scientifica|
|Titolo:||The minimal active domain of the mouse Ras exchange factor CDC25Mm|
|Autori:||Coccetti, P; Mauri, I; Alberghina, L; Martegani, E; Parmeggiani, A|
|Data di pubblicazione:||1995|
|Rivista:||BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1006/bbrc.1995.1035|
|Appare nelle tipologie:||01 - Articolo su rivista|