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Your search keyword '"M, Zerial"' showing total 17 results
Start Over Author "M, Zerial" Journal methods in enzymology
17 results on '"M, Zerial"'

2. Expression, purification, and characterization of Rab5 effector complex, rabaptin-5/rabex-5

Author

R, Lippé, H, Horiuchi, A, Runge, and M, Zerial

Subjects
Nitrilotriacetic Acid, Macromolecular Substances, Phosphatidylethanolamines, Genetic Vectors, Vesicular Transport Proteins, Brain, Membrane Proteins, Spodoptera, Chromatography, Affinity, Recombinant Proteins, Cytosol, Chromatography, Gel, Animals, Guanine Nucleotide Exchange Factors, Cattle, Electrophoresis, Polyacrylamide Gel, Baculoviridae, Protein Binding, rab5 GTP-Binding Proteins
Published
2001

3. Using oligonucleotides for cloning of Rab proteins by polymerase chain reaction

Author

L, Martín-Parras and M, Zerial

Subjects
DNA, Complementary, Base Sequence, GTP-Binding Proteins, Molecular Sequence Data, Mutagenesis, Site-Directed, Animals, RNA-Directed DNA Polymerase, Amino Acid Sequence, Cloning, Molecular, Codon, Polymerase Chain Reaction, DNA Primers, GTP Phosphohydrolases
Published
1995

4. Purification of posttranslationally modified and unmodified Rab5 protein expressed in Spodoptera frugiperda cells

Author

H, Horiuchi, O, Ullrich, C, Bucci, and M, Zerial

Subjects
Cell Membrane, Spodoptera, Cell Fractionation, Sulfur Radioisotopes, Transfection, Recombinant Proteins, Cell Line, Cytosol, GTP-Binding Proteins, Guanosine 5'-O-(3-Thiotriphosphate), Animals, Autoradiography, Electrophoresis, Polyacrylamide Gel, Baculoviridae, Protein Processing, Post-Translational, rab5 GTP-Binding Proteins
Published
1995

5. Use of Rab-GDP dissociation inhibitor for solubilization and delivery of Rab proteins to biological membranes in streptolysin O-permeabilized cells

Author

O, Ullrich, H, Horiuchi, K, Alexandrov, and M, Zerial

Subjects
Radioisotope Dilution Technique, Cell Membrane Permeability, Restriction Mapping, Kidney, Sulfur Radioisotopes, Tritium, Guanosine Diphosphate, Cell Line, GTP Phosphohydrolases, Dogs, Bacterial Proteins, GTP-Binding Proteins, Culture Techniques, Escherichia coli, Tumor Cells, Cultured, Animals, Cloning, Molecular, Guanine Nucleotide Dissociation Inhibitors, Sequence Tagged Sites, Cell Membrane, Chromatography, Ion Exchange, Recombinant Proteins, Kinetics, Solubility, Guanosine 5'-O-(3-Thiotriphosphate), Streptolysins, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel
Published
1995

6. Expression of Rab proteins during mouse embryonic development

Author

C, Murphy and M, Zerial

Subjects
Histological Techniques, Molecular Sequence Data, Uterus, Muscle, Smooth, Embryo, Mammalian, Kidney, Immunohistochemistry, Antibodies, Peptide Fragments, GTP Phosphohydrolases, Embryonic and Fetal Development, Mice, Gene Expression Regulation, GTP-Binding Proteins, Paraffin, Pregnancy, Intestine, Small, Animals, Female, Indicators and Reagents, Amino Acid Sequence, Rabbits
Published
1995

7. Expression of Rab GTPases using recombinant vaccinia viruses

Author

H, Stenmark, C, Bucci, and M, Zerial

Subjects
Recombination, Genetic, Genetic Vectors, Fluorescent Antibody Technique, Gene Expression, Vaccinia virus, Kidney, Transfection, Recombinant Proteins, Cell Line, GTP Phosphohydrolases, Kinetics, Microscopy, Fluorescence, GTP-Binding Proteins, Cricetinae, Liposomes, Receptors, Transferrin, Animals, Humans
Published
1995

8. Localization of Rab family members in animal cells

Author

M, Zerial, R, Parton, P, Chavrier, and R, Frank

Subjects
Molecular Sequence Data, Fluorescent Antibody Technique, Gene Expression, Biological Transport, Vaccinia virus, DNA-Directed RNA Polymerases, Transfection, Recombinant Proteins, Cell Compartmentation, Genes, ras, GTP-Binding Proteins, Bacteriophage T7, Animals, Amino Acid Sequence, Cloning, Molecular, Microscopy, Immunoelectron, Gene Library
Published
1992

9. Regulation of endosome dynamics by Rab5 and Huntingtin-HAP40 effector complex in physiological versus pathological conditions.

Author

Pal A, Severin F, Höpfner S, and Zerial M

Subjects
Actins metabolism, Animals, Blotting, Western, Cell Movement drug effects, Endocytosis physiology, Endosomes metabolism, HeLa Cells, Humans, Huntingtin Protein, Intracellular Signaling Peptides and Proteins, Mice, Microtubules metabolism, Nerve Tissue Proteins physiology, Nuclear Proteins physiology, Protein Transport, Carrier Proteins physiology, Endocytosis drug effects, rab5 GTP-Binding Proteins physiology
Abstract

Vesicular transport of signaling molecules, specifically neurotrophins, in neurons is essential for their differentiation, survival, and plasticity. Neurotrophins such as neuron growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are internalized by receptor-mediated endocytosis at synaptic terminals and loaded into endosomes for microtubule-based transport along axons to the cell body where they exert their signaling function in the nucleus. The molecular mechanisms underlying this intracellular transport are not only relevant from a basic knowledge viewpoint, but have also important implications for neurodegenerative diseases. Defects in trafficking are increasingly implicated in the pathology of Huntington's disease (HD) and other neurodegenerative disorders. The small GTPases Rab5 and Rab7 play important roles in the endocytic trafficking of neurotrophins. We have recently identified Huntingtin (Htt) and Huntingtin associated protein of 40 kDa (HAP40) as a novel Rab5 effector complex that regulates endosome motility. In HD, we detected higher HAP40 protein levels compared with normal cells. Such increase causes an augmented recruitment of Htt onto Rab5-positive early endosomes that drastically reduces their motility by "switching" these organelles from microtubules to F-actin. These findings suggest a mechanism by which impaired Rab5-mediated trafficking of neurotrophic factors may be a key event of the pathogenetic process leading to neurodegeneration in HD. To dissect the mechanisms by which Htt, HAP40, and Rab5 function in early endosome interactions with the cytoskeleton, we developed assays to investigate endosome-cytoskeleton interactions that can be applied to normal and pathological conditions. We provide here detailed protocols for, first, an assay that measures binding of early endosomes to microtubules and F-actin. Second, we describe an improved protocol for a cell-free assay that recapitulates the motility of early endosomes along microtubules in vitro. These assays provide mechanistic insights into the dysfunction of endosome motility occurring in HD as well as other neurodegenerative disorders.

Published
2008
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10. Expression, purification, and characterization of Rab5 effector complex, rabaptin-5/rabex-5.

Author

Lippé R, Horiuchi H, Runge A, and Zerial M

Subjects
Animals, Baculoviridae genetics, Brain cytology, Brain metabolism, Cattle, Chromatography, Affinity methods, Chromatography, Gel, Cytosol chemistry, Electrophoresis, Polyacrylamide Gel, Genetic Vectors genetics, Guanine Nucleotide Exchange Factors genetics, Macromolecular Substances, Membrane Proteins genetics, Phosphatidylethanolamines, Protein Binding, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, Spodoptera, rab5 GTP-Binding Proteins isolation & purification, Guanine Nucleotide Exchange Factors isolation & purification, Guanine Nucleotide Exchange Factors metabolism, Membrane Proteins isolation & purification, Membrane Proteins metabolism, Nitrilotriacetic Acid analogs & derivatives, Vesicular Transport Proteins, rab5 GTP-Binding Proteins metabolism
Published
2001
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11. Purification of EEA1 from bovine brain cytosol using Rab5 affinity chromatography and activity assays.

Author

Christoforidis S and Zerial M

Subjects
Animals, Brain cytology, Cattle, Chromatography, Gel methods, Chromatography, Ion Exchange methods, Electrophoresis, Polyacrylamide Gel, Transfection, Vesicular Transport Proteins, rab5 GTP-Binding Proteins genetics, Brain Chemistry, Chromatography, Affinity methods, Cytosol chemistry, Membrane Proteins isolation & purification, Membrane Proteins metabolism, rab5 GTP-Binding Proteins metabolism
Published
2001
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13. Purification of posttranslationally modified and unmodified Rab5 protein expressed in Spodoptera frugiperda cells.

Author

Horiuchi H, Ullrich O, Bucci C, and Zerial M

Subjects
Animals, Autoradiography methods, Baculoviridae, Cell Fractionation methods, Cell Line, Cell Membrane metabolism, Cytosol metabolism, Electrophoresis, Polyacrylamide Gel methods, GTP-Binding Proteins metabolism, Guanosine 5'-O-(3-Thiotriphosphate) metabolism, Recombinant Proteins biosynthesis, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, Spodoptera, Sulfur Radioisotopes, Transfection methods, rab5 GTP-Binding Proteins, GTP-Binding Proteins biosynthesis, GTP-Binding Proteins isolation & purification, Protein Processing, Post-Translational
Published
1995
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14. Expression of Rab proteins during mouse embryonic development.

Author

Murphy C and Zerial M

Subjects
Amino Acid Sequence, Animals, Antibodies, Embryo, Mammalian cytology, Female, GTP Phosphohydrolases analysis, GTP-Binding Proteins analysis, Histological Techniques, Immunohistochemistry methods, Indicators and Reagents, Intestine, Small cytology, Intestine, Small embryology, Intestine, Small ultrastructure, Kidney cytology, Kidney embryology, Kidney ultrastructure, Mice, Molecular Sequence Data, Muscle, Smooth cytology, Muscle, Smooth embryology, Muscle, Smooth ultrastructure, Paraffin, Peptide Fragments chemical synthesis, Peptide Fragments immunology, Pregnancy, Rabbits immunology, Uterus cytology, Uterus physiology, Embryo, Mammalian physiology, Embryonic and Fetal Development, GTP Phosphohydrolases biosynthesis, GTP-Binding Proteins biosynthesis, Gene Expression Regulation
Published
1995
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15. Use of Rab-GDP dissociation inhibitor for solubilization and delivery of Rab proteins to biological membranes in streptolysin O-permeabilized cells.

Author

Ullrich O, Horiuchi H, Alexandrov K, and Zerial M

Subjects
Animals, Bacterial Proteins, Cell Line, Cell Membrane metabolism, Cell Membrane Permeability, Chromatography, Gel methods, Chromatography, Ion Exchange methods, Cloning, Molecular methods, Culture Techniques methods, Dogs, Electrophoresis, Polyacrylamide Gel methods, Escherichia coli, GTP Phosphohydrolases analysis, GTP-Binding Proteins analysis, Guanosine 5'-O-(3-Thiotriphosphate) metabolism, Guanosine Diphosphate metabolism, Kidney, Kinetics, Radioisotope Dilution Technique, Recombinant Proteins analysis, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, Restriction Mapping, Sequence Tagged Sites, Solubility, Streptolysins, Sulfur Radioisotopes, Tritium, Tumor Cells, Cultured, GTP Phosphohydrolases isolation & purification, GTP Phosphohydrolases metabolism, GTP-Binding Proteins isolation & purification, GTP-Binding Proteins metabolism, Guanine Nucleotide Dissociation Inhibitors
Published
1995
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16. Expression of Rab GTPases using recombinant vaccinia viruses.

Author

Stenmark H, Bucci C, and Zerial M

Subjects
Animals, Cell Line, Cricetinae, Fluorescent Antibody Technique, GTP Phosphohydrolases analysis, GTP-Binding Proteins analysis, Gene Expression, Genetic Vectors, Humans, Kidney, Kinetics, Liposomes, Microscopy, Fluorescence methods, Receptors, Transferrin analysis, Receptors, Transferrin biosynthesis, Recombinant Proteins analysis, Recombination, Genetic, Transfection methods, Vaccinia virus, GTP Phosphohydrolases biosynthesis, GTP-Binding Proteins biosynthesis, Recombinant Proteins biosynthesis
Published
1995
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17. Localization of Rab family members in animal cells.

Author

Zerial M, Parton R, Chavrier P, and Frank R

Subjects
Amino Acid Sequence, Animals, Bacteriophage T7, Biological Transport, Cell Compartmentation, Cloning, Molecular, DNA-Directed RNA Polymerases, Fluorescent Antibody Technique, GTP-Binding Proteins biosynthesis, GTP-Binding Proteins genetics, GTP-Binding Proteins immunology, Gene Expression, Gene Library, Microscopy, Immunoelectron, Molecular Sequence Data, Recombinant Proteins immunology, Recombinant Proteins isolation & purification, Transfection, Vaccinia virus genetics, GTP-Binding Proteins isolation & purification, Genes, ras genetics, Recombinant Proteins biosynthesis
Published
1992
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