Identification of Human Cadherin-14, a Novel Neurally Specific Type II Cadherin, by Protein Interaction Cloning

Shibata, Tatsuhiro; Shimoyama, Yutaka; Gotoh, Masahiro; Hirohashi, Setsuo

doi:10.1074/jbc.272.8.5236

Cited by 44 publications

(39 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Northern blotting analysis was performed as described previously. 24 Ten micrograms of total RNA from the cell lines or 2 g of messenger RNA (mRNA) from multiple human tissues (Clontech) were electrophoresed and hybridized with 32 P-radiolabeled EBP50, GAPDH, and ␤-actin cDNA.…”

Section: Methodsmentioning

confidence: 99%

“…In vitro protein interaction screening and proteinbinding assay were performed as described. 24 A fragment of human ␤-catenin complementary DNA (cDNA) (amino acids 350-781), which contains 7 armadillo repeats and a carboxyl domain, was subcloned into pGEX2TK (Amersham Pharmacia, Buckinghamshire, United Kingdom). Recombinant glutathione S-transferase (GST) fusion protein was purified with glutathione sepharose (Amersham Pharmacia) and labeled with 32 P-␥-ATP and bovine heart protein kinase (Sigma, St. Louis, MO).…”

Section: Methodsmentioning

confidence: 99%

“…Immunoprecipitation and immunoblotting were performed with mouse monoclonal anti-FLAG M2 antibody (Sigma), rabbit polyclonal anti-␤-catenin antibody (Chemicon, Temecula, CA), mouse monoclonal anti-E-cadherin antibody (Transduction Laboratory, Lexington, KY), mouse monoclonal anti-EBP50 antibody (Transduction Laboratory), and mouse monoclonal anti-GFP antibody (Clontech) as described. 24 For in vitro protein-binding assay, the PDZ-1 and PDZ-2 fragments of EBP50 or the wild type and mutated carboxyl domain of ␤-catenin were amplified by PCR, subcloned in pGEX4T (Amersham Pharmacia). Recombinant proteins (10 g) conjugated with glutathione sepharose were mixed with cellular protein lysed in lysis buffer (10 mmol/L, Tris, pH 7.5, 150 mmol/L NaCl, 0.5% Triton X-100, and protease inhibitor mix [Roche, Mannheim, Germany]).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

EBP50, a β-catenin-associating protein, enhances Wnt signaling and is over-expressed in hepatocellular carcinoma

Shibata¹

2003

Hepatology

146

View full text Add to dashboard Cite

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

EBP50, a β-catenin-associating protein, enhances Wnt signaling and is over-expressed in hepatocellular carcinoma

Shibata¹

2003

Hepatology

146

View full text Add to dashboard Cite

“…cDNA Cloning-To use ␤-catenin as a probe for the first cDNA cloning procedure based on the protein-protein interaction, we constructed and purified ␤-catenin-glutathione S-transferase fusion protein, as described previously (39). This protein was radiolabeled in vitro with [␥-32 P]ATP using bovine heart kinase and was used to screen a human adult brain gt11 expression cDNA library (CLONTECH).…”

Section: Methodsmentioning

confidence: 99%

“…By using this method, we found two novel human classic cadherins, which we named cadherin-14 and -15. The former is a novel type II classic cadherin expressed widely in the central nervous system, and recently, we reported its cDNA cloning (39), and the latter closely resembles mouse M (muscle)-cadherin and appears to be its human counterpart. However, this molecule has an additional peptide sequence at its C terminus not possessed by M-cadherin.…”

mentioning

confidence: 99%

Molecular Cloning and Characterization of a Novel Human Classic Cadherin Homologous with Mouse Muscle Cadherin

Shimoyama¹,

Shibata²,

Kitajima³

et al. 1998

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

We used a novel cDNA cloning method based on the cadherin-␤-catenin protein interaction and identified a new human classic-type cadherin, which we named cadherin-15, from adult brain and skeletal muscle cDNA libraries. Sequence analysis revealed that this cadherin was closely related to mouse muscle cadherin and seemed to be its human counterpart. However, its deduced amino acid sequence differed from that of mouse muscle cadherin in that it had an extra 31-amino acid sequence at its C terminus that has been found neither in mouse muscle cadherin nor in any other known classic cadherin. Analysis of cadherin-15 protein expressed in L fibroblasts showed that it was cleaved proteolytically, expressed on the cell surfaces as a mature form of about 124-kDa, and functioned as a cell-cell adhesion molecule in a homophilic and specific manner, but Ca 2؉ did not protect it against degradation by trypsin. Our findings also suggest that cadherin-15 mediates cell-cell adhesion with a binding strength comparable to that of E-cadherin.It is generally accepted that various molecules that have the extracellular subdomain (EC) 1 structure of the classic cadherins in common constitute a large gene family, the cadherin superfamily. This superfamily includes the classic cadherins (1), truncated-type cadherins, which lack the characteristic cytoplasmic domain of the classic cadherins (2-4), desmosomal cadherins, which are localized in desmosomes (5-7), protocadherins, which have more than five extracellular subdomains (8 -9), and molecules showing high similarities to rat LI-cadherin (10 -12).Each classic cadherin comprises a signal sequence and a precursor region, which are both cleaved by intracellular proteolytic processing, five cadherin extracellular subdomain repeats, a transmembrane domain and a characteristic cytoplasmic domain, which is highly conserved among the subclasses and is indispensable for association with catenins, the ensuing linkage to the cytoskeleton and full functioning as a Ca 2ϩ -dependent cell-cell adhesion molecule (13-15). It is now understood that these cadherins play essential roles in various morphogenetic events in multicellular organisms (1). The first classic cadherins to be identified were E-, N-, and P-cadherins, as a result of the establishment of their respective blocking antibodies (16 -23), and then V-cadherin was identified using a blocking monoclonal antibody (24). Thereafter, several cadherins were identified (25-28) by determining their cross-reactivities with antibodies raised against conserved peptide sequences or cross-hybridization with cDNA fragments of known cadherins. Over the past few years, the existence of more classic cadherin molecules has been demonstrated by PCR-based cDNA cloning methods (3, 29 -34).Full cDNA cloning of nine independent human classic cadherin molecules has been reported, i.e. E-, N-, and P-cadherins and cadherin-4, -5, -6, -8, -11 (OB-cadherin), and -12 (3, 29, 30, 31, 35-37). We are interested in how many classic cadherin molecules actually exist in humans...

show abstract

Germline Copy Number Variation and Cancer Risk

Voer

Venkatachalam

Kessel

et al. 2012

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Cancer is usually considered to be an acquired disease caused by the progressive accumulation of somatic deoxyribonucleic acid (DNA) changes. It is clear, however, that inherited factors may also play a significant role in the initiation of this disease. Some of these factors represent loss‐of‐function mutations in tumour suppressor genes, resulting in an increased cancer risk among carriers. Such an increased risk may also be attributed to the presence of multiple polymorphic variants such as single nucleotide polymorphisms (SNPs), each of which may convey a mild effect upon cancer susceptibility. DNA copy number variation (CNV) and other structural variations in the human genome are increasingly recognised as an alternative source of genetic variation that may influence cancer risk. Specifically, rare CNVs may affect important cancer‐associated genes and/or pathways and, thus, provide an explanation for moderate‐ to high‐risk cancer families. Therefore, it is anticipated that the identification of rare germline CNVs in unexplained familial and early onset cancer patients will contribute to our understanding of cancer predisposition and development. Key Concepts: Copy number variation (CNV) is a common form of normal variation affecting a significant proportion of the human genome. CNVs frequently encompass annotated genes. Disruption and/or silencing of critical genes located in rare CNVs may result in increased risk of disease, including cancer. Genome‐wide copy number analysis can be used as a strategy to identify novel moderate‐ to high‐penetrant cancer predisposing genes and/or mechanisms. Discovery of cancer‐predisposing CNVs may reveal new cancer syndromes that exhibit additional clinical features.

show abstract

Identification of Human Cadherin-14, a Novel Neurally Specific Type II Cadherin, by Protein Interaction Cloning

Cited by 44 publications

References 40 publications

EBP50, a β-catenin-associating protein, enhances Wnt signaling and is over-expressed in hepatocellular carcinoma

EBP50, a β-catenin-associating protein, enhances Wnt signaling and is over-expressed in hepatocellular carcinoma

Molecular Cloning and Characterization of a Novel Human Classic Cadherin Homologous with Mouse Muscle Cadherin

Germline Copy Number Variation and Cancer Risk

Contact Info

Product

Resources

About