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Clonning and characterization of the membrane microdomain targeted componets
dc.contributor.advisorČerný, Jan
dc.creatorFalteisek, Lukáš
dc.date.accessioned2017-04-12T15:46:49Z
dc.date.available2017-04-12T15:46:49Z
dc.date.issued2008
dc.identifier.urihttp://hdl.handle.net/20.500.11956/1282
dc.description.abstract3 Abstract Lipid rafts are defined as islets in the membranes of eucaryotic cells with different composition from the rest of the membrane. They appear spontaneously due to phase separation of different membrane components and are proposed to serve as platforms for concentration of selected signaling proteins. However, evidence for their existence is still indirect, despite more than decade of intense research. Some new approaches show that the fluctuations of membrane composition are more diverse and are caused more likely by presence of proteins than by lipid phase separation. We performed bioinformatical search looking for new signaling proteins targeted to putative rafts. We have identified several proteins and out of them phosphodiesterase 8a (PDE 8a) was salected for further research. To prepare the mouse monoclonal antibody we expressed and purified fragment of PDE 8a fused with GST. After immunisation we obtained one clone producing antibodies tentatively positive on western blot and by indirect immunofluorescence. We predicted that PDE 8a is targeted to putative rafts by N-terminal myristoylation and palmitoylation. To clarify, whether these modifications are present in the strusture of PDE 8a we prepared mutants of the PDE 8a N-terminal region lacking myristoylation, palmitoylation or both. These...en_US
dc.languageČeštinacs_CZ
dc.language.isocs_CZ
dc.publisherUniverzita Karlova, Přírodovědecká fakultacs_CZ
dc.titleKlonování a charakterizace proteinů potenciálně zacílených do membránových mikrodoméncs_CZ
dc.typediplomová prácecs_CZ
dcterms.created2008
dcterms.dateAccepted2008-09-22
dc.description.departmentKatedra fyziol. živočichů a vývoj. biol. (zrušena)cs_CZ
dc.description.departmentDep. of Physiology and Develop. Biology (obsolete)en_US
dc.description.facultyFaculty of Scienceen_US
dc.description.facultyPřírodovědecká fakultacs_CZ
dc.identifier.repId33041
dc.title.translatedClonning and characterization of the membrane microdomain targeted componetsen_US
dc.contributor.refereeBrábek, Jan
dc.identifier.aleph001008195
thesis.degree.nameMgr.
thesis.degree.levelnavazující magisterskécs_CZ
thesis.degree.disciplineImunologiecs_CZ
thesis.degree.disciplineImmunologyen_US
thesis.degree.programBiologiecs_CZ
thesis.degree.programBiologyen_US
uk.thesis.typediplomová prácecs_CZ
uk.taxonomy.organization-csPřírodovědecká fakulta::Katedra fyziol. živočichů a vývoj. biol. (zrušena)cs_CZ
uk.taxonomy.organization-enFaculty of Science::Dep. of Physiology and Develop. Biology (obsolete)en_US
uk.faculty-name.csPřírodovědecká fakultacs_CZ
uk.faculty-name.enFaculty of Scienceen_US
uk.faculty-abbr.csPřFcs_CZ
uk.degree-discipline.csImunologiecs_CZ
uk.degree-discipline.enImmunologyen_US
uk.degree-program.csBiologiecs_CZ
uk.degree-program.enBiologyen_US
thesis.grade.csVýborněcs_CZ
thesis.grade.enExcellenten_US
uk.abstract.en3 Abstract Lipid rafts are defined as islets in the membranes of eucaryotic cells with different composition from the rest of the membrane. They appear spontaneously due to phase separation of different membrane components and are proposed to serve as platforms for concentration of selected signaling proteins. However, evidence for their existence is still indirect, despite more than decade of intense research. Some new approaches show that the fluctuations of membrane composition are more diverse and are caused more likely by presence of proteins than by lipid phase separation. We performed bioinformatical search looking for new signaling proteins targeted to putative rafts. We have identified several proteins and out of them phosphodiesterase 8a (PDE 8a) was salected for further research. To prepare the mouse monoclonal antibody we expressed and purified fragment of PDE 8a fused with GST. After immunisation we obtained one clone producing antibodies tentatively positive on western blot and by indirect immunofluorescence. We predicted that PDE 8a is targeted to putative rafts by N-terminal myristoylation and palmitoylation. To clarify, whether these modifications are present in the strusture of PDE 8a we prepared mutants of the PDE 8a N-terminal region lacking myristoylation, palmitoylation or both. These...en_US
uk.file-availabilityV
uk.publication.placePrahacs_CZ
uk.grantorUniverzita Karlova, Přírodovědecká fakulta, Katedra fyziol. živočichů a vývoj. biol. (zrušena)cs_CZ
dc.identifier.lisID990010081950106986


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