The functional domain of the stimulatory GDP/GTP exchange protein (smg GDS) which interacts with the C-terminal geranylgeranylated region of rap1/Krev-1/smg p21.
rap1/Krev-1/smg p21 (smg p21), a member of the small GTP-binding protein (G protein) superfamily, has a geranylgeranylated cysteine residue and clustered basic amino acids in the C-terminal region. The GDP/GTP exchange reaction of smg p21 is regulated by smg GDS, which is also active on Ki-ras p21 and rho p21. The C-terminal region of smg p21 is essential for its interaction with smg GDS. Moreover, smg p21 is phosphorylated by cyclic AMP- and cyclic GMP-dependent protein kinases at the serine residue between the polybasic region and the prenylated cysteine residue, and this phosphorylation initiates the smg GDS-induced smg p21 activation. Thus, the C-terminal cationic and hydrophobic region is important for the regulation of the smg p21 activity. In the present study, we attempted to determine the functional domain of smg GDS which interacts with the C-terminal region of smg p21 by use of a cross-link method and a site-directed mutagenesis method. The region of smg GDS cross-linked with the C-terminal region of smg p21B was residues 444-492, which is located at the C-terminal fifth of smg GDS. On deletion of these residues, smg GDS became inactive on smg p21B, Ki-ras p21 and rhoA p21. These results indicate that residues 444-492 of smg GDS are at least one of the domains which interact with the C-terminal region of its substrate small G proteins.
Kotani
K
K
Department of Biochemistry, Kobe University, School of Medicine, Japan.
Kikuchi
A
A
Doi
K
K
Kishida
S
S
Sakoda
T
T
Kishi
K
K
Takai
Y
Y
D000595:Amino Acid Sequence; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011506:Proteins / Q000378:metabolism*; D013329:Structure-Activity Relationship; D020668:rap GTP-Binding Proteins
D011506:Proteins; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D020668:rap GTP-Binding Proteins