TY - JOUR
T1 - Identification of a novel β-catenin-interacting protein
AU - Kawajiri, Aie
AU - Itoh, Naohiro
AU - Fukata, Masaki
AU - Nakagawa, Masato
AU - Yamaga, Masaki
AU - Iwamatsu, Akihiro
AU - Kaibuchi, Kozo
N1 - Funding Information:
Abbreviations used: GEF, guanine nucleotide exchange factor; GST, glutathione S-transferase; aa, amino acid; SDS–PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; PBS, phosphate-buffered saline; MDCKII, Madin–Darby canine kidney II. 1This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, and Culture of Japan (1999) and by grants from the program Research for the Future of the Japan Society for the Promotion of Science, the Human Frontier Science Program, and Kirin Brewery Company Limited. 2These authors contributed equally to this article. 3To whom correspondence should be addressed at Division of Signal Transduction, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, 630-0101, Japan. Fax: 0743-72-5449. E-mail: [email protected].
PY - 2000/7/5
Y1 - 2000/7/5
N2 - Cadherin is a well-known cell-cell adhesion molecule, and it binds to β-catenin, which in turn binds to α-catenin. However, little is known about the regulatory mechanism underlying the cadherin-mediated cell-cell adhesion. Here we purified two novel β-catenin-interacting proteins with molecular masses of 180 kDa (p180) and 150 kDa (p150) from bovine brain cytosol by using glutathione S-transferase (GST)-β-catenin affinity column chromatography. Mass spectral analysis revealed p180 to be identical to KIAA0313 which has a putative Rap1 guanine nucleotide exchange factor (GEF) domain and p150 to be the same as KIAA0705 which has a high degree of sequence similarity to the synaptic scaffolding molecule (S-SCAM), which binds β-catenin and KIAA0313 in the yeast two-hybrid system and overlay assay, respectively. β-Catenin was coimmunoprecipitated with KIAA0313 in Madin-Darby canine kidney II (MDCKII) cells, bovine brain cytosol, and EL cells. KIAA0313 and β-catenin were partly colocalized at sites of cell-cell contact in MDCKII cells. Taken together, our data suggest that KIAA0313 associates with β-catenin through KIAA0705 in vivo at sites of cell-cell contact. (C) 2000 Academic Press.
AB - Cadherin is a well-known cell-cell adhesion molecule, and it binds to β-catenin, which in turn binds to α-catenin. However, little is known about the regulatory mechanism underlying the cadherin-mediated cell-cell adhesion. Here we purified two novel β-catenin-interacting proteins with molecular masses of 180 kDa (p180) and 150 kDa (p150) from bovine brain cytosol by using glutathione S-transferase (GST)-β-catenin affinity column chromatography. Mass spectral analysis revealed p180 to be identical to KIAA0313 which has a putative Rap1 guanine nucleotide exchange factor (GEF) domain and p150 to be the same as KIAA0705 which has a high degree of sequence similarity to the synaptic scaffolding molecule (S-SCAM), which binds β-catenin and KIAA0313 in the yeast two-hybrid system and overlay assay, respectively. β-Catenin was coimmunoprecipitated with KIAA0313 in Madin-Darby canine kidney II (MDCKII) cells, bovine brain cytosol, and EL cells. KIAA0313 and β-catenin were partly colocalized at sites of cell-cell contact in MDCKII cells. Taken together, our data suggest that KIAA0313 associates with β-catenin through KIAA0705 in vivo at sites of cell-cell contact. (C) 2000 Academic Press.
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U2 - 10.1006/bbrc.2000.3002
DO - 10.1006/bbrc.2000.3002
M3 - Article
C2 - 10873669
AN - SCOPUS:0033873750
SN - 0006-291X
VL - 273
SP - 712
EP - 717
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -