{"id":{"repo_id":"helsinki","oai_identifier":"oai:helda.helsinki.fi:10138/591384"},"canonical_url":"https://search.dev.ndltd.org/etd/helsinki/oai:helda.helsinki.fi:10138/591384","repository":{"repo_id":"helsinki","name":"University of Helsinki","base_url":"https://helda.helsinki.fi/server/oai/request"},"display":{"title":"Protein interaction studies on the titin A150 domain using proximity-dependent biotinylation","abstract":"Titin is an enormous protein that spans over 1 µm in the sarcomere, making it the largest protein in the human body. A single missense variant within the titin A150 domain, also called FN3 119, is sufficient to cause a dominant myopathy known as Hereditary Myopathy with Early Respiratory Failure (HMERF), with the most prevalent disease-causing variant being the c.95134T>C (p.C31712R) variant. HMERF is characterized by weakness in proximal and distal skeletal muscles and severe, life-threatening, respiratory insufficiency. Despite its clinical significance, the pathomechanism behind the disease remains unknown. The aim of this thesis was to screen for protein-protein interactions of both the wild-type titin A150 domain and the A150 domain containing the disease-causing variant p.C31712R, with the goal of gaining deeper insight into the protein interactions of titin and the molecular mechanism behind HMERF. In this study, proximity dependent biotinylation (PDB) was used to screen for protein interactions. PDB was conducted using the biotin-ligase UltraID fused to a titin fragment. This study encompasses the creation of the titin-UltraID fusion genes, the functional validation of the titin UltraID proteins, the development of a PDB workflow and the analysis of identified proteins. The study was conducted in stable, differentiated and electrostimulated C2C12 myotubes grown on a gelatin hydrogel. Although the proteins identified were not statistically significant, this thesis provides insight into potential interactors of the titin A150 domain and the A150 domain with the pathogenic p.C31712R variant. This thesis also offers insight into the use of PDB for studying titinopathies in a C2C12 cell model. However, further validation is needed to fully substantiate the findings in this thesis.","abstract_html":"Titin is an enormous protein that spans over 1 µm in the sarcomere, making it the largest protein in the human body. A single missense variant within the titin A150 domain, also called FN3 119, is sufficient to cause a dominant myopathy known as Hereditary Myopathy with Early Respiratory Failure (HMERF), with the most prevalent disease-causing variant being the c.95134T>C (p.C31712R) variant. HMERF is characterized by weakness in proximal and distal skeletal muscles and severe, life-threatening, respiratory insufficiency. Despite its clinical significance, the pathomechanism behind the disease remains unknown. The aim of this thesis was to screen for protein-protein interactions of both the wild-type titin A150 domain and the A150 domain containing the disease-causing variant p.C31712R, with the goal of gaining deeper insight into the protein interactions of titin and the molecular mechanism behind HMERF. In this study, proximity dependent biotinylation (PDB) was used to screen for protein interactions. PDB was conducted using the biotin-ligase UltraID fused to a titin fragment. This study encompasses the creation of the titin-UltraID fusion genes, the functional validation of the titin UltraID proteins, the development of a PDB workflow and the analysis of identified proteins. The study was conducted in stable, differentiated and electrostimulated C2C12 myotubes grown on a gelatin hydrogel. Although the proteins identified were not statistically significant, this thesis provides insight into potential interactors of the titin A150 domain and the A150 domain with the pathogenic p.C31712R variant. This thesis also offers insight into the use of PDB for studying titinopathies in a C2C12 cell model. However, further validation is needed to fully substantiate the findings in this thesis.","abstract_has_math":false,"creators":["Vainio, Anna Linnéa"],"institution":"Helsingin yliopisto","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-01-29","date_published":"2025-01-29","updated_at":"2026-07-27T19:56:01Z","subjects":["titin","titinopathy","Hereditary myopathy with early respiratory failure","HMERF","proximity dependent biotinylation","protein interactomics","UltraID","BioID","biotin labelling","proximity labelling","myopathy"],"languages":["eng"],"rights":["CC BY-NC-ND 4.0"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10138/591384","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Vainio, Anna Linnéa"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2025-01-29T09:27:18Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2025-01-29T09:27:18Z"]},{"key":"dc:date.issued","label":"Date","values":["2025-01-29"]},{"key":"dc:publisher","label":"Institution","values":["Helsingin yliopisto","University of Helsinki","Helsingfors universitet"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["titin","titinopathy","Hereditary myopathy with early respiratory failure","HMERF","proximity dependent biotinylation","protein interactomics","UltraID","BioID","biotin labelling","proximity labelling","myopathy"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["CC BY-NC-ND 4.0"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/10138/591384"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Titin is an enormous protein that spans over 1 µm in the sarcomere, making it the largest protein in the human body. A single missense variant within the titin A150 domain, also called FN3 119, is sufficient to cause a dominant myopathy known as Hereditary Myopathy with Early Respiratory Failure (HMERF), with the most prevalent disease-causing variant being the c.95134T>C (p.C31712R) variant. HMERF is characterized by weakness in proximal and distal skeletal muscles and severe, life-threatening, respiratory insufficiency. Despite its clinical significance, the pathomechanism behind the disease remains unknown. The aim of this thesis was to screen for protein-protein interactions of both the wild-type titin A150 domain and the A150 domain containing the disease-causing variant p.C31712R, with the goal of gaining deeper insight into the protein interactions of titin and the molecular mechanism behind HMERF. In this study, proximity dependent biotinylation (PDB) was used to screen for protein interactions. PDB was conducted using the biotin-ligase UltraID fused to a titin fragment. This study encompasses the creation of the titin-UltraID fusion genes, the functional validation of the titin UltraID proteins, the development of a PDB workflow and the analysis of identified proteins. The study was conducted in stable, differentiated and electrostimulated C2C12 myotubes grown on a gelatin hydrogel. Although the proteins identified were not statistically significant, this thesis provides insight into potential interactors of the titin A150 domain and the A150 domain with the pathogenic p.C31712R variant. This thesis also offers insight into the use of PDB for studying titinopathies in a C2C12 cell model. However, further validation is needed to fully substantiate the findings in this thesis.","Titin sträcker över 1 µm i sarkomeren och är därmed det största proteinet i människokroppen. Flera olika missense-varianter i titinets A150 domän, även kallad FN3 119, har kopplats till den dominanta muskelsjukdomen HMERF (eng. Hereditary Myopathy with Early Respiratory Failure) varav den mest förekommande sjukdomsalstrande varianten är punktmutationen c.95134T>C (p.C31712R). Symptomerna för HMERF innefattar bland annat svaghet i proximala och distala skelettmuskler samt livshotande andningssvikt. Trots sjukdomens allvarliga kliniska manifestationer är de molekylära mekanismerna bakom sjukdomen fortfarande oklara. Syftet med denna magisteravhandling var att undersöka protein-protein interaktioner av både vildtyp titinets A150 domän och det patogena A150 domänet med p.C31712R missense-varianten, för att få insikter i domänets proteininteraktioner samt för att få bättre förståelse av de molekylära mekanismerna bakom HMERF. I denna studie användes proximitetsberoende biotinmärkning (eng. proximity-dependent biotinylation (PDB)) för att undersöka proteininteraktioner. PDB genomfördes via användningen av titin fragment försett med biotin-ligaset UltraID. Denna avhandling innefattar skapandet av titin-UltraID fusion gener, testandet av titin-UltraID proteinernas funktionalitet, utvecklandet av ett PDB protokoll och analys av identifierade proteiner. Studien var gjord i stabila, differentierade och elektrostimulerade C2C12 myotuber. Fastän de identifierade proteinerna inte var statistiskt signifikanta ger denna avhandling insikter i potentiella proteininteraktorer för både viltyp A150 domänet och domänet med patogena varianten. Denna avhandling ger också insikter i användningen av PDB för att studera titin i en C2C12 cell modell. Ytterligare validering behövs dock för att bekräfta resultaten i denna avhandling."]},{"key":"dc:title","label":"Title","values":["Protein interaction studies on the titin A150 domain using proximity-dependent biotinylation"]}]}],"canonical_facts":{"dc:creator":["Vainio, Anna Linnéa"],"dc:date.accessioned":["2025-01-29T09:27:18Z"],"dc:date.available":["2025-01-29T09:27:18Z"],"dc:date.issued":["2025-01-29"],"dc:description.abstract":["Titin is an enormous protein that spans over 1 µm in the sarcomere, making it the largest protein in the human body. A single missense variant within the titin A150 domain, also called FN3 119, is sufficient to cause a dominant myopathy known as Hereditary Myopathy with Early Respiratory Failure (HMERF), with the most prevalent disease-causing variant being the c.95134T>C (p.C31712R) variant. HMERF is characterized by weakness in proximal and distal skeletal muscles and severe, life-threatening, respiratory insufficiency. Despite its clinical significance, the pathomechanism behind the disease remains unknown. The aim of this thesis was to screen for protein-protein interactions of both the wild-type titin A150 domain and the A150 domain containing the disease-causing variant p.C31712R, with the goal of gaining deeper insight into the protein interactions of titin and the molecular mechanism behind HMERF. In this study, proximity dependent biotinylation (PDB) was used to screen for protein interactions. PDB was conducted using the biotin-ligase UltraID fused to a titin fragment. This study encompasses the creation of the titin-UltraID fusion genes, the functional validation of the titin UltraID proteins, the development of a PDB workflow and the analysis of identified proteins. The study was conducted in stable, differentiated and electrostimulated C2C12 myotubes grown on a gelatin hydrogel. Although the proteins identified were not statistically significant, this thesis provides insight into potential interactors of the titin A150 domain and the A150 domain with the pathogenic p.C31712R variant. This thesis also offers insight into the use of PDB for studying titinopathies in a C2C12 cell model. However, further validation is needed to fully substantiate the findings in this thesis.","Titin sträcker över 1 µm i sarkomeren och är därmed det största proteinet i människokroppen. Flera olika missense-varianter i titinets A150 domän, även kallad FN3 119, har kopplats till den dominanta muskelsjukdomen HMERF (eng. Hereditary Myopathy with Early Respiratory Failure) varav den mest förekommande sjukdomsalstrande varianten är punktmutationen c.95134T>C (p.C31712R). Symptomerna för HMERF innefattar bland annat svaghet i proximala och distala skelettmuskler samt livshotande andningssvikt. Trots sjukdomens allvarliga kliniska manifestationer är de molekylära mekanismerna bakom sjukdomen fortfarande oklara. Syftet med denna magisteravhandling var att undersöka protein-protein interaktioner av både vildtyp titinets A150 domän och det patogena A150 domänet med p.C31712R missense-varianten, för att få insikter i domänets proteininteraktioner samt för att få bättre förståelse av de molekylära mekanismerna bakom HMERF. I denna studie användes proximitetsberoende biotinmärkning (eng. proximity-dependent biotinylation (PDB)) för att undersöka proteininteraktioner. PDB genomfördes via användningen av titin fragment försett med biotin-ligaset UltraID. Denna avhandling innefattar skapandet av titin-UltraID fusion gener, testandet av titin-UltraID proteinernas funktionalitet, utvecklandet av ett PDB protokoll och analys av identifierade proteiner. Studien var gjord i stabila, differentierade och elektrostimulerade C2C12 myotuber. Fastän de identifierade proteinerna inte var statistiskt signifikanta ger denna avhandling insikter i potentiella proteininteraktorer för både viltyp A150 domänet och domänet med patogena varianten. Denna avhandling ger också insikter i användningen av PDB för att studera titin i en C2C12 cell modell. Ytterligare validering behövs dock för att bekräfta resultaten i denna avhandling."],"dc:identifier.uri":["http://hdl.handle.net/10138/591384"],"dc:language.iso":["eng"],"dc:publisher":["Helsingin yliopisto","University of Helsinki","Helsingfors universitet"],"dc:rights":["CC BY-NC-ND 4.0"],"dc:subject":["titin","titinopathy","Hereditary myopathy with early respiratory failure","HMERF","proximity dependent biotinylation","protein interactomics","UltraID","BioID","biotin labelling","proximity labelling","myopathy"],"dc:title":["Protein interaction studies on the titin A150 domain using proximity-dependent biotinylation"]},"updated_at":"2026-07-27T19:56:01Z"}