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Endosomal escape of canine parvovirus is assisted by membrane fluidization
S86
Abstracts / Chemistry and Physics of Lipids 149S (2007) S74–S91
PO 146
Transcriptome teleonomy and scavenger lipidomics of albumin and alpha 1-acid glycoproteins: contextual genome-wide ontology enrichment as evidence for the epigenetic code? Pauli Ojala 1 , Sami Kilpinen 2 , Lotta Arola 1 , Petri T¨or¨onen 3 , Elmar Bucher 2 , Kalle Ojala 2 , Kristiina Iljin 2 , Mari Peltonen 2 , Rolf Skotheim 2 , Kristine Kleivi 2 , Martti Tolvanen 4 , Reija Autio 5 , Pentti Somerharju 6 , Olli Kallioniemi 2 , Saara Laitinen 1 1 Blood
Service, Red Cross, Finland; 2 Medical Biotechnology, VTT Technical Research Centre of Finland and University of Turku, Finland; 3 Institute of Biotechnology, University of Helsinki, Finland; 4 Institute of Medical Technology, University of Tampere, Finland; 5 Institute of Signal Processing, Tampere University of Technology, Finland; 6 Department of Biochemistry, Institute of Biomedicine, University of Helsinki, Finland Orosomucoid (a-1-acid glycoprotein, AGP) is the most abundant plasma lipocalin (25–100 mcM), and a major acute phase marker of inflammation and infection from blood and urine with its 45% (w/w) glycans. AGP is a potent cytoprotectant e.g. in ischemic-reperfusion injury. We show ex vivo by ESI-MS and in vitro in equilibrium and kinetically by native and pyrene-labelled lipids that AGP scavenges LPC more tightly than HSA (140–180 nM Kd). AGP is less vulnerable than HSA to competition by fatty acids. Expression pattern of the proteins is analyzed from a database of 10,000 human Affymetrix microarrays (Kilpinen et al., unpublished). By collective normalization of the raw data (Autio et al., unpublished), extrahepatic tissue profiles are reported in high resolution. Threshold-free global enrichment of the metacorrelates against 17,500 human genes and 120 tissues is predicted as teleonomic, functional, purpose. Principal component analysis displays AGP1 and -2 as the closest coworkers in the genome, but very indifferent to HSA expression. The physical and causal literature relations of the metacorrelates are quantitated and visualized using the Ariadne Genomics Pathway Studio 5.0 and Resnet 5.0 knowledgebase. The induced AGP coexpression pattern encompasses Defense & Immune response, Glycoprotein, Glycosaminoglycan binding, Transendothelial migration, Signal transduction, Apoptosis, Cytokine-receptor network, Plasma, Acute phase response, and Lectin. On the other hand, homeostatic processes are specifically repressed. AGPs closest associates include bactericidal permeability-increasing
protein, formyl peptide receptor-like 1, haptoglobin, proteoglycan 2, natural killer cell activator, calgranulin C and B, PAF-receptor, and arachidonate 5-lipoxygenaseactivating protein. Thus, circulating AGP seems to regulate the innate immunity through a more specific binding and targeting of the inflammatory LPC than the FFA sequestering HSA. doi:10.1016/j.chemphyslip.2007.06.197 PO 147
Endosomal escape of canine parvovirus is assisted by membrane fluidization Kirsi Pakkanen, Anna R. M¨akel¨a, Sanna Kirjavainen, Katja S¨aa¨ j¨arvi, Nina Rintanen, Tuula O. Jalonen, Christian Oker-Blom, Matti Vuento Department of Biology and Environmental Science, University of Jyv¨askyl¨a, Finland Canine parvovirus (CPV) is a small, non-enveloped virus of the autonomous Parvovirus genus of Parvoviridae family. It enters its host cells by clathrin-mediated endocytosis via transferrin receptor and is trafficked to late stages of endocytic route, where it is released to cytoplasm. Although details of this release process are so far unclear, it is known that the phospholipase A2 activity in capsid protein VP1 is involved in membrane penetration. We studied the effect of canine parvovirus capsids to model and biological membranes using a naphtyl styryl dye di-4-ANEPPDHQ which reacts to changes in membrane fluidity with shifts in fluorescence emission maxima. According to our results, CPV induces an increase in the fluidity of rigid model membranes at pH 5.5 but not in neutral conditions. The effect was not seen with more fluid model membranes. Studies with a CPV permissive cell line, NLFK, have shown that increase in membrane fluidity occurs also during normal CPV infection on the endosomal membranes. In relation to this, our latest results show, that CPV is able to escape from endosomal vesicles even when the membranes of the cell are made more rigid with excess cholesterol or sphingomyelin accumulation. However, when in addition to sphingomyelin accumulation ceramide is fed to cells, CPV is retained in endosomal vesicles. These findings indicate that, in addition to the classical degradation functions of PLA2, also complex fluidity-modulating mechanisms are used in the vesicle release process of CPV. doi:10.1016/j.chemphyslip.2007.06.198
Abstracts / Chemistry and Physics of Lipids 149S (2007) S74–S91
PO 146
Transcriptome teleonomy and scavenger lipidomics of albumin and alpha 1-acid glycoproteins: contextual genome-wide ontology enrichment as evidence for the epigenetic code? Pauli Ojala 1 , Sami Kilpinen 2 , Lotta Arola 1 , Petri T¨or¨onen 3 , Elmar Bucher 2 , Kalle Ojala 2 , Kristiina Iljin 2 , Mari Peltonen 2 , Rolf Skotheim 2 , Kristine Kleivi 2 , Martti Tolvanen 4 , Reija Autio 5 , Pentti Somerharju 6 , Olli Kallioniemi 2 , Saara Laitinen 1 1 Blood
Service, Red Cross, Finland; 2 Medical Biotechnology, VTT Technical Research Centre of Finland and University of Turku, Finland; 3 Institute of Biotechnology, University of Helsinki, Finland; 4 Institute of Medical Technology, University of Tampere, Finland; 5 Institute of Signal Processing, Tampere University of Technology, Finland; 6 Department of Biochemistry, Institute of Biomedicine, University of Helsinki, Finland Orosomucoid (a-1-acid glycoprotein, AGP) is the most abundant plasma lipocalin (25–100 mcM), and a major acute phase marker of inflammation and infection from blood and urine with its 45% (w/w) glycans. AGP is a potent cytoprotectant e.g. in ischemic-reperfusion injury. We show ex vivo by ESI-MS and in vitro in equilibrium and kinetically by native and pyrene-labelled lipids that AGP scavenges LPC more tightly than HSA (140–180 nM Kd). AGP is less vulnerable than HSA to competition by fatty acids. Expression pattern of the proteins is analyzed from a database of 10,000 human Affymetrix microarrays (Kilpinen et al., unpublished). By collective normalization of the raw data (Autio et al., unpublished), extrahepatic tissue profiles are reported in high resolution. Threshold-free global enrichment of the metacorrelates against 17,500 human genes and 120 tissues is predicted as teleonomic, functional, purpose. Principal component analysis displays AGP1 and -2 as the closest coworkers in the genome, but very indifferent to HSA expression. The physical and causal literature relations of the metacorrelates are quantitated and visualized using the Ariadne Genomics Pathway Studio 5.0 and Resnet 5.0 knowledgebase. The induced AGP coexpression pattern encompasses Defense & Immune response, Glycoprotein, Glycosaminoglycan binding, Transendothelial migration, Signal transduction, Apoptosis, Cytokine-receptor network, Plasma, Acute phase response, and Lectin. On the other hand, homeostatic processes are specifically repressed. AGPs closest associates include bactericidal permeability-increasing
protein, formyl peptide receptor-like 1, haptoglobin, proteoglycan 2, natural killer cell activator, calgranulin C and B, PAF-receptor, and arachidonate 5-lipoxygenaseactivating protein. Thus, circulating AGP seems to regulate the innate immunity through a more specific binding and targeting of the inflammatory LPC than the FFA sequestering HSA. doi:10.1016/j.chemphyslip.2007.06.197 PO 147
Endosomal escape of canine parvovirus is assisted by membrane fluidization Kirsi Pakkanen, Anna R. M¨akel¨a, Sanna Kirjavainen, Katja S¨aa¨ j¨arvi, Nina Rintanen, Tuula O. Jalonen, Christian Oker-Blom, Matti Vuento Department of Biology and Environmental Science, University of Jyv¨askyl¨a, Finland Canine parvovirus (CPV) is a small, non-enveloped virus of the autonomous Parvovirus genus of Parvoviridae family. It enters its host cells by clathrin-mediated endocytosis via transferrin receptor and is trafficked to late stages of endocytic route, where it is released to cytoplasm. Although details of this release process are so far unclear, it is known that the phospholipase A2 activity in capsid protein VP1 is involved in membrane penetration. We studied the effect of canine parvovirus capsids to model and biological membranes using a naphtyl styryl dye di-4-ANEPPDHQ which reacts to changes in membrane fluidity with shifts in fluorescence emission maxima. According to our results, CPV induces an increase in the fluidity of rigid model membranes at pH 5.5 but not in neutral conditions. The effect was not seen with more fluid model membranes. Studies with a CPV permissive cell line, NLFK, have shown that increase in membrane fluidity occurs also during normal CPV infection on the endosomal membranes. In relation to this, our latest results show, that CPV is able to escape from endosomal vesicles even when the membranes of the cell are made more rigid with excess cholesterol or sphingomyelin accumulation. However, when in addition to sphingomyelin accumulation ceramide is fed to cells, CPV is retained in endosomal vesicles. These findings indicate that, in addition to the classical degradation functions of PLA2, also complex fluidity-modulating mechanisms are used in the vesicle release process of CPV. doi:10.1016/j.chemphyslip.2007.06.198