{"id":{"repo_id":"heid-diss","oai_identifier":"oai:archiv.ub.uni-heidelberg.de:3639"},"canonical_url":"https://search.dev.ndltd.org/etd/heid-diss/oai:archiv.ub.uni-heidelberg.de:3639","repository":{"repo_id":"heid-diss","name":"Universität Heidelberg","base_url":"http://archiv.ub.uni-heidelberg.de/volltextserver/cgi/oai2"},"display":{"title":"Structural and Functional Characterization of GAPR-1, a Mammalian Plant Pathogenesis-related Protein in Lipid-enriched Microdomains of the Golgi Complex","abstract":"During the characterization of lipid-enriched microdomains at the Golgi (GICs) (Gkantiragas, I. et al. 2001), a protein with an apparent molecular mass of 17 kDa was identified. Cloning and preliminary biochemical characterization identified a novel protein, GAPR-1, belonging to the superfamily of PR proteins. Based on the primary amino acidic sequence of this protein, some potentially interesting characteristics were identified. It contains a consensus sequence for myristoylation, a putative caveolin-binding domain, a coiled-coil structure, and an isolelectric point (pI) of 9.4, suggesting that GAPR-1 is a highly hydrophilic protein (Eberle, H. B. et al. 2002). In this thesis, this structural information, was used to i) study the interaction of GAPR-1 with membranes, ii) to obtain structural information on the protein, and iii) to identify proteins that interact with GAPR-1.GAPR-1 was shown to be myristoylated and to interact with Caveolin-1. Myristoylation, together with protein-protein or electrostatic interactions at physiological pH could explain its strong membrane association. The crystal structure of GAPR-1 showed strong structural similarities to other plant pathogenesis-related proteins. Substitution of the most conserved amino acids in GAPR-1 (His54, Glu65, Glu86 and His103) in the putative active center changed the protein behavior in solution. Size exclusion chromatography revealed that the major population of GAPR-1 mutant migrated as a dimer, whereas GAPR-1 wild type behaves predominantly as a monomer. The tendency of GAPR-1 to form dimers was confirmed by crosslink experiments and by the yeast two hybrid system. By affinity chromatography, GAPR-1 was shown to interact with three proteins: Nucleolin, Template activating factor α (TAFIα) and HSAPRIL. In the yeast two hybrid system, the interaction of GARP-1 with Nucleolin was confirmed and shown to be dependent on the most conserved amino acid residues in GAPR-1. The interaction between GAPR-1 and Nucleolin may represent a new mechanism of regulation of innate immunity in mammalian cells.","abstract_html":"During the characterization of lipid-enriched microdomains at the Golgi (GICs) (Gkantiragas, I. et al. 2001), a protein with an apparent molecular mass of 17 kDa was identified. Cloning and preliminary biochemical characterization identified a novel protein, GAPR-1, belonging to the superfamily of PR proteins. Based on the primary amino acidic sequence of this protein, some potentially interesting characteristics were identified. It contains a consensus sequence for myristoylation, a putative caveolin-binding domain, a coiled-coil structure, and an isolelectric point (pI) of 9.4, suggesting that GAPR-1 is a highly hydrophilic protein (Eberle, H. B. et al. 2002). In this thesis, this structural information, was used to i) study the interaction of GAPR-1 with membranes, ii) to obtain structural information on the protein, and iii) to identify proteins that interact with GAPR-1.GAPR-1 was shown to be myristoylated and to interact with Caveolin-1. Myristoylation, together with protein-protein or electrostatic interactions at physiological pH could explain its strong membrane association. The crystal structure of GAPR-1 showed strong structural similarities to other plant pathogenesis-related proteins. Substitution of the most conserved amino acids in GAPR-1 (His54, Glu65, Glu86 and His103) in the putative active center changed the protein behavior in solution. Size exclusion chromatography revealed that the major population of GAPR-1 mutant migrated as a dimer, whereas GAPR-1 wild type behaves predominantly as a monomer. The tendency of GAPR-1 to form dimers was confirmed by crosslink experiments and by the yeast two hybrid system. By affinity chromatography, GAPR-1 was shown to interact with three proteins: Nucleolin, Template activating factor α (TAFIα) and HSAPRIL. In the yeast two hybrid system, the interaction of GARP-1 with Nucleolin was confirmed and shown to be dependent on the most conserved amino acid residues in GAPR-1. The interaction between GAPR-1 and Nucleolin may represent a new mechanism of regulation of innate immunity in mammalian cells.","abstract_has_math":false,"creators":["Serrano, Ramon Leonardo"],"institution":"Universität Heidelberg","degree_name":null,"degree_level":"thesis.doctoral","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Wieland, Felix Professor"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2003,"date_issued":"2003-07-09","date_published":"2003-07-09","updated_at":"2026-07-24T02:29:34Z","subjects":[],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://www.ub.uni-heidelberg.de/archiv/3639","outbound_label":"Repository record","outbound_source":"source_url"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Wieland, Felix Professor"]},{"key":"dc:creator","label":"Author","values":["Serrano, Ramon Leonardo"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:publisher","label":"Institution","values":["Universitätsbibliothek Heidelberg"]},{"key":"dc:type","label":"Dc Type","values":["doctoralThesis"]},{"key":"thesis:degree_level","label":"Degree Level","values":["thesis.doctoral"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Universität Heidelberg"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["During the characterization of lipid-enriched microdomains at the Golgi (GICs) (Gkantiragas, I. et al. 2001), a protein with an apparent molecular mass of 17 kDa was identified. Cloning and preliminary biochemical characterization identified a novel protein, GAPR-1, belonging to the superfamily of PR proteins. Based on the primary amino acidic sequence of this protein, some potentially interesting characteristics were identified. It contains a consensus sequence for myristoylation, a putative caveolin-binding domain, a coiled-coil structure, and an isolelectric point (pI) of 9.4, suggesting that GAPR-1 is a highly hydrophilic protein (Eberle, H. B. et al. 2002). In this thesis, this structural information, was used to i) study the interaction of GAPR-1 with membranes, ii) to obtain structural information on the protein, and iii) to identify proteins that interact with GAPR-1.GAPR-1 was shown to be myristoylated and to interact with Caveolin-1. Myristoylation, together with protein-protein or electrostatic interactions at physiological pH could explain its strong membrane association. The crystal structure of GAPR-1 showed strong structural similarities to other plant pathogenesis-related proteins. Substitution of the most conserved amino acids in GAPR-1 (His54, Glu65, Glu86 and His103) in the putative active center changed the protein behavior in solution. Size exclusion chromatography revealed that the major population of GAPR-1 mutant migrated as a dimer, whereas GAPR-1 wild type behaves predominantly as a monomer. The tendency of GAPR-1 to form dimers was confirmed by crosslink experiments and by the yeast two hybrid system. By affinity chromatography, GAPR-1 was shown to interact with three proteins: Nucleolin, Template activating factor α (TAFIα) and HSAPRIL. In the yeast two hybrid system, the interaction of GARP-1 with Nucleolin was confirmed and shown to be dependent on the most conserved amino acid residues in GAPR-1. The interaction between GAPR-1 and Nucleolin may represent a new mechanism of regulation of innate immunity in mammalian cells."]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Structural and Functional Characterization of GAPR-1, a Mammalian Plant Pathogenesis-related Protein in Lipid-enriched Microdomains of the Golgi Complex"]}]}],"canonical_facts":{"dc:contributor":["Wieland, Felix Professor"],"dc:creator":["Serrano, Ramon Leonardo"],"dc:description.abstract":["During the characterization of lipid-enriched microdomains at the Golgi (GICs) (Gkantiragas, I. et al. 2001), a protein with an apparent molecular mass of 17 kDa was identified. Cloning and preliminary biochemical characterization identified a novel protein, GAPR-1, belonging to the superfamily of PR proteins. Based on the primary amino acidic sequence of this protein, some potentially interesting characteristics were identified. It contains a consensus sequence for myristoylation, a putative caveolin-binding domain, a coiled-coil structure, and an isolelectric point (pI) of 9.4, suggesting that GAPR-1 is a highly hydrophilic protein (Eberle, H. B. et al. 2002). In this thesis, this structural information, was used to i) study the interaction of GAPR-1 with membranes, ii) to obtain structural information on the protein, and iii) to identify proteins that interact with GAPR-1.GAPR-1 was shown to be myristoylated and to interact with Caveolin-1. Myristoylation, together with protein-protein or electrostatic interactions at physiological pH could explain its strong membrane association. The crystal structure of GAPR-1 showed strong structural similarities to other plant pathogenesis-related proteins. Substitution of the most conserved amino acids in GAPR-1 (His54, Glu65, Glu86 and His103) in the putative active center changed the protein behavior in solution. Size exclusion chromatography revealed that the major population of GAPR-1 mutant migrated as a dimer, whereas GAPR-1 wild type behaves predominantly as a monomer. The tendency of GAPR-1 to form dimers was confirmed by crosslink experiments and by the yeast two hybrid system. By affinity chromatography, GAPR-1 was shown to interact with three proteins: Nucleolin, Template activating factor α (TAFIα) and HSAPRIL. In the yeast two hybrid system, the interaction of GARP-1 with Nucleolin was confirmed and shown to be dependent on the most conserved amino acid residues in GAPR-1. The interaction between GAPR-1 and Nucleolin may represent a new mechanism of regulation of innate immunity in mammalian cells."],"dc:format.medium":["application/pdf"],"dc:publisher":["Universitätsbibliothek Heidelberg"],"dc:title":["Structural and Functional Characterization of GAPR-1, a Mammalian Plant Pathogenesis-related Protein in Lipid-enriched Microdomains of the Golgi Complex"],"dc:type":["doctoralThesis"],"thesis:degree_level":["thesis.doctoral"],"thesis:institution_name":["Universität Heidelberg"]},"updated_at":"2026-07-24T02:29:34Z"}