{"id":{"repo_id":"aachen","oai_identifier":"oai:publications.rwth-aachen.de:56238"},"canonical_url":"https://search.dev.ndltd.org/etd/aachen/oai:publications.rwth-aachen.de:56238","repository":{"repo_id":"aachen","name":"RWTH Aachen University","base_url":"https://publications.rwth-aachen.de/oai2d"},"display":{"title":"Proteomics based analysis of interactions between CEA cell adhesion molecule 1 (CEACAM1) and intracellular proteins in transfected tumor cells","abstract":"CEA Cell Adhesion Molecule 1 (CEACAM1), a type 1 transmembrane protein belonging to the CEA gene family, is alternatively spliced to produce four major isoforms with a long (CEACAM1-L) or a short cytoplasmic domain (CEACAM1-S). Immunoprecipitations using the murine adenocarcinoma MC38 cell line transfected with human CEACAM1-L were performed to isolate cytoplasmic proteins binding to the unphosphorylated or tyrosine phosphorylated CEACAM1 long cytoplasmic domain. Mass spectrometric analysis of immunoprecipitates separated on two-dimensional gels revealed actin, tropomyosin, myosin, vimentin and cytokeratin as the major proteins interacting with CEACAM1-L after sodium pervanadate induced tyrosine phosphorylation of CEACAM1-L. Surface plasmon resonance studies were performed to characterize actin and tropomyosin binding to the membrane-proximate region of the CEACAM1-L cytoplasmic domain and to its short splicing variant CEACAM1-S. A CEACAM1-L derived peptide bound F-actin and tropomyosin with KDs of 1.3x10–5 M, and 1.8x10-5 M, respectively, while an equivalent CEACAM1-S peptide bound less than 10% of the long form to F-actin and tropomyosin with a KD <10-4 M. GST-long or short cytoplasmic domain fusion proteins bound actin and tropomyosin with KDs of 3.0x10-8 M and 4.1x10-7 M, respectively, for the long form, and KDs of 6.2x10-8 M and 2.6x10-7 M, respectively, for the short form. Calmodulin, a known CEACAM1 binding protein, or EDTA inhibited binding of the CEACAM1-L peptide and both GST-CEACAM1 fusion proteins to actin, while calmodulin and actin, but not EDTA, stimulated binding of GST-CEACAM1-L to tropomyosin. Confocal microscopy studies showed a sodium pervanadate dependent colocalization of F-actin and CEACAM1-L at cell-cell adhesion sites in CEACAM1-L transfected MC38 cells, confirming the initial immunoprecipitation results. The identification of an association of both CEACAM1 cytoplasmic domains with actin and tropomyosin and its potential regulation by calmodulin and Ca2+ could lead to a better understanding of the mechanisms and the functions of interactions between transmembrane cell adhesion proteins and the cytoskeleton.","abstract_html":"CEA Cell Adhesion Molecule 1 (CEACAM1), a type 1 transmembrane protein belonging to the CEA gene family, is alternatively spliced to produce four major isoforms with a long (CEACAM1-L) or a short cytoplasmic domain (CEACAM1-S). Immunoprecipitations using the murine adenocarcinoma MC38 cell line transfected with human CEACAM1-L were performed to isolate cytoplasmic proteins binding to the unphosphorylated or tyrosine phosphorylated CEACAM1 long cytoplasmic domain. Mass spectrometric analysis of immunoprecipitates separated on two-dimensional gels revealed actin, tropomyosin, myosin, vimentin and cytokeratin as the major proteins interacting with CEACAM1-L after sodium pervanadate induced tyrosine phosphorylation of CEACAM1-L. Surface plasmon resonance studies were performed to characterize actin and tropomyosin binding to the membrane-proximate region of the CEACAM1-L cytoplasmic domain and to its short splicing variant CEACAM1-S. A CEACAM1-L derived peptide bound F-actin and tropomyosin with KDs of 1.3x10–5 M, and 1.8x10-5 M, respectively, while an equivalent CEACAM1-S peptide bound less than 10% of the long form to F-actin and tropomyosin with a KD &lt;10-4 M. GST-long or short cytoplasmic domain fusion proteins bound actin and tropomyosin with KDs of 3.0x10-8 M and 4.1x10-7 M, respectively, for the long form, and KDs of 6.2x10-8 M and 2.6x10-7 M, respectively, for the short form. Calmodulin, a known CEACAM1 binding protein, or EDTA inhibited binding of the CEACAM1-L peptide and both GST-CEACAM1 fusion proteins to actin, while calmodulin and actin, but not EDTA, stimulated binding of GST-CEACAM1-L to tropomyosin. Confocal microscopy studies showed a sodium pervanadate dependent colocalization of F-actin and CEACAM1-L at cell-cell adhesion sites in CEACAM1-L transfected MC38 cells, confirming the initial immunoprecipitation results. The identification of an association of both CEACAM1 cytoplasmic domains with actin and tropomyosin and its potential regulation by calmodulin and Ca2+ could lead to a better understanding of the mechanisms and the functions of interactions between transmembrane cell adhesion proteins and the cytoskeleton.","abstract_has_math":false,"creators":["Schumann, Detlef"],"institution":"Publikationsserver der RWTH Aachen University","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Kreuzaler, Fritz"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2000,"date_issued":"2000","date_published":"2000","updated_at":"2026-07-30T19:41:53Z","subjects":["info:eu-repo/classification/ddc/570","Biowissenschaften, Biologie","Carcino-embryonales Antigen","Zell-Adhäsionsmolekül","Wechselwirkung","Tumorzelle","Intrazellulärraum","Proteine"],"languages":["eng"],"rights":["info:eu-repo/semantics/openAccess"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-118356%22"],"render_values":[{"text":"https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-118356%22","href":"https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-118356%22","code":true}]}]},"links":{"outbound_url":"https://publications.rwth-aachen.de/record/56238","outbound_label":"Repository record","outbound_source":"dc:identifier"},"source_record":{"url":"https://publications.rwth-aachen.de/oai2d?verb=GetRecord&metadataPrefix=oai_dc&identifier=oai%3Apublications.rwth-aachen.de%3A56238","prefix":"oai_dc"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Kreuzaler, Fritz"]},{"key":"dc:creator","label":"Author","values":["Schumann, Detlef"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:coverage","label":"Dc Coverage","values":["DE"]},{"key":"dc:date","label":"Dc Date","values":["2000"]},{"key":"dc:publisher","label":"Institution","values":["Publikationsserver der RWTH Aachen University"]},{"key":"dc:relation","label":"Dc Relation","values":["info:eu-repo/semantics/altIdentifier/urn/urn:nbn:de:hbz:82-opus-367"]},{"key":"dc:type","label":"Dc Type","values":["info:eu-repo/semantics/doctoralThesis","info:eu-repo/semantics/publishedVersion"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["info:eu-repo/classification/ddc/570","Biowissenschaften, Biologie","Carcino-embryonales Antigen","Zell-Adhäsionsmolekül","Wechselwirkung","Tumorzelle","Intrazellulärraum","Proteine"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["info:eu-repo/semantics/openAccess"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://publications.rwth-aachen.de/record/56238","https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-118356%22"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["CEA Cell Adhesion Molecule 1 (CEACAM1), a type 1 transmembrane protein belonging to the CEA gene family, is alternatively spliced to produce four major isoforms with a long (CEACAM1-L) or a short cytoplasmic domain (CEACAM1-S). Immunoprecipitations using the murine adenocarcinoma MC38 cell line transfected with human CEACAM1-L were performed to isolate cytoplasmic proteins binding to the unphosphorylated or tyrosine phosphorylated CEACAM1 long cytoplasmic domain. Mass spectrometric analysis of immunoprecipitates separated on two-dimensional gels revealed actin, tropomyosin, myosin, vimentin and cytokeratin as the major proteins interacting with CEACAM1-L after sodium pervanadate induced tyrosine phosphorylation of CEACAM1-L. Surface plasmon resonance studies were performed to characterize actin and tropomyosin binding to the membrane-proximate region of the CEACAM1-L cytoplasmic domain and to its short splicing variant CEACAM1-S. A CEACAM1-L derived peptide bound F-actin and tropomyosin with KDs of 1.3x10–5 M, and 1.8x10-5 M, respectively, while an equivalent CEACAM1-S peptide bound less than 10% of the long form to F-actin and tropomyosin with a KD <10-4 M. GST-long or short cytoplasmic domain fusion proteins bound actin and tropomyosin with KDs of 3.0x10-8 M and 4.1x10-7 M, respectively, for the long form, and KDs of 6.2x10-8 M and 2.6x10-7 M, respectively, for the short form. Calmodulin, a known CEACAM1 binding protein, or EDTA inhibited binding of the CEACAM1-L peptide and both GST-CEACAM1 fusion proteins to actin, while calmodulin and actin, but not EDTA, stimulated binding of GST-CEACAM1-L to tropomyosin. Confocal microscopy studies showed a sodium pervanadate dependent colocalization of F-actin and CEACAM1-L at cell-cell adhesion sites in CEACAM1-L transfected MC38 cells, confirming the initial immunoprecipitation results. The identification of an association of both CEACAM1 cytoplasmic domains with actin and tropomyosin and its potential regulation by calmodulin and Ca2+ could lead to a better understanding of the mechanisms and the functions of interactions between transmembrane cell adhesion proteins and the cytoskeleton."]},{"key":"dc:source","label":"Dc Source","values":["Aachen : Publikationsserver der RWTH Aachen University III, 105 S. : graph. Darst. (2000). = Aachen, Techn. Hochsch., Diss., 2000"]},{"key":"dc:title","label":"Title","values":["Proteomics based analysis of interactions between CEA cell adhesion molecule 1 (CEACAM1) and intracellular proteins in transfected tumor cells"]}]}],"canonical_facts":{"dc:contributor":["Kreuzaler, Fritz"],"dc:coverage":["DE"],"dc:creator":["Schumann, Detlef"],"dc:date":["2000"],"dc:description":["CEA Cell Adhesion Molecule 1 (CEACAM1), a type 1 transmembrane protein belonging to the CEA gene family, is alternatively spliced to produce four major isoforms with a long (CEACAM1-L) or a short cytoplasmic domain (CEACAM1-S). Immunoprecipitations using the murine adenocarcinoma MC38 cell line transfected with human CEACAM1-L were performed to isolate cytoplasmic proteins binding to the unphosphorylated or tyrosine phosphorylated CEACAM1 long cytoplasmic domain. Mass spectrometric analysis of immunoprecipitates separated on two-dimensional gels revealed actin, tropomyosin, myosin, vimentin and cytokeratin as the major proteins interacting with CEACAM1-L after sodium pervanadate induced tyrosine phosphorylation of CEACAM1-L. Surface plasmon resonance studies were performed to characterize actin and tropomyosin binding to the membrane-proximate region of the CEACAM1-L cytoplasmic domain and to its short splicing variant CEACAM1-S. A CEACAM1-L derived peptide bound F-actin and tropomyosin with KDs of 1.3x10–5 M, and 1.8x10-5 M, respectively, while an equivalent CEACAM1-S peptide bound less than 10% of the long form to F-actin and tropomyosin with a KD <10-4 M. GST-long or short cytoplasmic domain fusion proteins bound actin and tropomyosin with KDs of 3.0x10-8 M and 4.1x10-7 M, respectively, for the long form, and KDs of 6.2x10-8 M and 2.6x10-7 M, respectively, for the short form. Calmodulin, a known CEACAM1 binding protein, or EDTA inhibited binding of the CEACAM1-L peptide and both GST-CEACAM1 fusion proteins to actin, while calmodulin and actin, but not EDTA, stimulated binding of GST-CEACAM1-L to tropomyosin. Confocal microscopy studies showed a sodium pervanadate dependent colocalization of F-actin and CEACAM1-L at cell-cell adhesion sites in CEACAM1-L transfected MC38 cells, confirming the initial immunoprecipitation results. The identification of an association of both CEACAM1 cytoplasmic domains with actin and tropomyosin and its potential regulation by calmodulin and Ca2+ could lead to a better understanding of the mechanisms and the functions of interactions between transmembrane cell adhesion proteins and the cytoskeleton."],"dc:identifier":["https://publications.rwth-aachen.de/record/56238","https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-118356%22"],"dc:language":["eng"],"dc:publisher":["Publikationsserver der RWTH Aachen University"],"dc:relation":["info:eu-repo/semantics/altIdentifier/urn/urn:nbn:de:hbz:82-opus-367"],"dc:rights":["info:eu-repo/semantics/openAccess"],"dc:source":["Aachen : Publikationsserver der RWTH Aachen University III, 105 S. : graph. Darst. (2000). = Aachen, Techn. Hochsch., Diss., 2000"],"dc:subject":["info:eu-repo/classification/ddc/570","Biowissenschaften, Biologie","Carcino-embryonales Antigen","Zell-Adhäsionsmolekül","Wechselwirkung","Tumorzelle","Intrazellulärraum","Proteine"],"dc:title":["Proteomics based analysis of interactions between CEA cell adhesion molecule 1 (CEACAM1) and intracellular proteins in transfected tumor cells"],"dc:type":["info:eu-repo/semantics/doctoralThesis","info:eu-repo/semantics/publishedVersion"]},"updated_at":"2026-07-30T19:41:53Z"}