Peroxisomes

Softcover Nachdruck of the original 1. Auflage 1994
ISBN: 978-3-642-87809-1
Verlag: Springer

Buch, Englisch, 201 Seiten, Paperback, Format (B × H): 140 mm x 216 mm, Gewicht: 283 g

Reihe: Springer Lab Manuals

Discovered and first isolated in 1966 in C. De Duve's
laboratory, peroxisomes - organelles which are present in
nearly all eukaryotic cells - are still not fully
understood.
More than 40 peroxisomal enzymes catalyzing a variety of
reactions have been characterized. Moreover, peroxisomes can
be regarded as toxicological indicators: several compounds,
including hypolipemic drugs, plasticizers or pesticides
trigger their proliferation. This proliferation may lead
to hepatocarcinogenesis in rodents.
Interest in peroxisomes stems not only fromtheir biology,
but also because there is a deficiency of peroxisomal
functions in several genetic diseases. Some genes involved
in inborn errors of peroxisomal function have recently been
identified.
In this manual, based on a FEBS Advanced Course on
peroxisomes, protocols on the following topics are described
in detail: Isolation and characterization of peroxisomes by
ultracentrifugationand immunoblotting; gene regulation
studied by mRNA isolation, hybridizationand DNA cell
transfection; use of cell lines as peroxisome proliferator
targets; transformation with retrovirus; peroxisomes as
toxicological markers; cytochrome P450 induction; drug
design and computer analysis of ligand/receptor interaction
involved in peroxisomal gene expression.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Bugaut, Maurice

Latruffe, Norbert

Fachgebiete

• Medizin | Veterinärmedizin Medizin | Public Health | Pharmazie | Zahnmedizin Medizinische Fachgebiete Pharmakologie, Toxikologie
• Naturwissenschaften Biowissenschaften Zellbiologie
• Naturwissenschaften Biowissenschaften Biochemie (nichtmedizinisch)
• Medizin | Veterinärmedizin Medizin | Public Health | Pharmazie | Zahnmedizin Klinische und Innere Medizin

Inhaltsverzeichnis

I: Isolation and Characterization of Peroxisomes.- General Indroduction to Isolation and Characterization of Peroxisomes.- Experiment 1. Preparation and Purification of Peroxisomes; Subfractionation of Purified Peroxisomes; Acyl-CoA Oxidase Activity Measurements.- Experiment 2. Immunoblotting of Peroxisomal Proteins with Monospecific Antibodies.- Experiment 3. Morphology of Peroxisomes in Light- and Electron Microscopy.- II: Molecular Biology.- Experiment 4. Northern Blotting Analysis of Rat Liver mRNA Encoding for Peroxisomal Acyl-CoA Oxidase.- Experiment 5. Transient Co-transfection Assay: Measure of the Chloramphenicol Acetyl Transferase (CAT) Activity in Cytosol Extracts from Transfected Cells.- Experiment 6. Synthesis of Oligonucleotides Used as Probes; Purification by HPLC.- Experiment 7. Computer Analysis of DNA and Protein Sequences.- III: Toxicology and Pharmacology.- Experiment 8. Cytochrome P450 Isoenzyme Induction Pattern in Rats Treated with Peroxisome Proliferators and Classical Inducers.- Experiment 9. Measurement of the in Vitro Glucuronidation of Peroxisome Proliferator Carboxylic Acids by Liver Microsomes and by Genetically Modified V 79 Cell Line.- Experiment 10. Molecular Modeling and Drug Design: Application to Some Peroxisome Proliferator Agents.- IV: Cell Culture and Genetic Diseases.- Experiment 11. Fao Cell Line as a Model for the Study of the Effect of Peroxisome Proliferators on Cellular Functions.- Experiment 12. Measurement of Dihydroxyacetone-Phosphate Acyl-Transferase (DHAP-AT) Activity in Liver Peroxisomes and Cell Lines.- Experiment 13. In Vitro Transformation of Human Fibroblasts with SV40 T Antigen (Lipofection).- Experiment 14. Somatic Cell Hybridization as a Tool for Genetic Analysis of Peroxisomal Activities.- Supplement.