The Experts below are selected from a list of 303 Experts worldwide ranked by ideXlab platform
David Macherel - One of the best experts on this subject based on the ideXlab platform.
-
Experimental determination of organelle targeting Peptide Cleavage sites using transient expression of GFP translational fusions
Analytical Biochemistry, 2013Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Helene Rogniaux-bonaventure, Marie-hélène Avelange-macherel, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with GFP fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles, and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting Peptide sequences and is suitable for animal and plant organelle-targeted proteins.
-
Experimental determination of organelle targeting-Peptide Cleavage sites using transient expression of green fluorescent protein translational fusions.
Analytical biochemistry, 2012Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Marie-hélène Avelange-macherel, Hélène Rogniaux, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence, which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with green fluorescent protein (GFP) fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting-Peptide sequences and is suitable for animal and plant organelle-targeted proteins.
-
Experimental determination of organelle targeting Peptide Cleavage sites using transient expression of GFP translational fusions
2012Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Helene Rogniaux-bonaventure, Marie-hélène Avelange-macherel, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence, which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with green fluorescent protein (GFP) fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting-Peptide sequences and is suitable for animal and plant organelle-targeted proteins.
László Patthy - One of the best experts on this subject based on the ideXlab platform.
-
Insertion of spliceosomal introns in proto-splice sites: the case of secretory signal Peptides.
FEBS letters, 2004Co-Authors: Hedvig Tordai, László PatthyAbstract:Analysis of the exon-intron structures of 2208 human genes has revealed that there is a statistically highly significant excess of phase 1 introns in the vicinity of the signal Peptide Cleavage sites. It is suggested that amino acid sequences surrounding signal Peptide Cleavage sites are significantly enriched in phase 1 proto-splice sites and this has favored insertion of spliceosomal introns in these sites.
-
Insertion of spliceosomal introns in proto‐splice sites: the case of secretory signal Peptides
FEBS Letters, 2004Co-Authors: Hedvig Tordai, László PatthyAbstract:Analysis of the exon–intron structures of 2208 human genes has revealed that there is a statistically highly significant excess of phase 1 introns in the vicinity of the signal Peptide Cleavage sites. It is suggested that amino acid sequences surrounding signal Peptide Cleavage sites are significantly enriched in phase 1 proto-splice sites and this has favored insertion of spliceosomal introns in these sites.
Adrien Candat - One of the best experts on this subject based on the ideXlab platform.
-
Experimental determination of organelle targeting Peptide Cleavage sites using transient expression of GFP translational fusions
Analytical Biochemistry, 2013Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Helene Rogniaux-bonaventure, Marie-hélène Avelange-macherel, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with GFP fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles, and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting Peptide sequences and is suitable for animal and plant organelle-targeted proteins.
-
Experimental determination of organelle targeting-Peptide Cleavage sites using transient expression of green fluorescent protein translational fusions.
Analytical biochemistry, 2012Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Marie-hélène Avelange-macherel, Hélène Rogniaux, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence, which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with green fluorescent protein (GFP) fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting-Peptide sequences and is suitable for animal and plant organelle-targeted proteins.
-
Experimental determination of organelle targeting Peptide Cleavage sites using transient expression of GFP translational fusions
2012Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Helene Rogniaux-bonaventure, Marie-hélène Avelange-macherel, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence, which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with green fluorescent protein (GFP) fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting-Peptide sequences and is suitable for animal and plant organelle-targeted proteins.
Hedvig Tordai - One of the best experts on this subject based on the ideXlab platform.
-
Insertion of spliceosomal introns in proto-splice sites: the case of secretory signal Peptides.
FEBS letters, 2004Co-Authors: Hedvig Tordai, László PatthyAbstract:Analysis of the exon-intron structures of 2208 human genes has revealed that there is a statistically highly significant excess of phase 1 introns in the vicinity of the signal Peptide Cleavage sites. It is suggested that amino acid sequences surrounding signal Peptide Cleavage sites are significantly enriched in phase 1 proto-splice sites and this has favored insertion of spliceosomal introns in these sites.
-
Insertion of spliceosomal introns in proto‐splice sites: the case of secretory signal Peptides
FEBS Letters, 2004Co-Authors: Hedvig Tordai, László PatthyAbstract:Analysis of the exon–intron structures of 2208 human genes has revealed that there is a statistically highly significant excess of phase 1 introns in the vicinity of the signal Peptide Cleavage sites. It is suggested that amino acid sequences surrounding signal Peptide Cleavage sites are significantly enriched in phase 1 proto-splice sites and this has favored insertion of spliceosomal introns in these sites.
Marie-hélène Avelange-macherel - One of the best experts on this subject based on the ideXlab platform.
-
Experimental determination of organelle targeting Peptide Cleavage sites using transient expression of GFP translational fusions
Analytical Biochemistry, 2013Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Helene Rogniaux-bonaventure, Marie-hélène Avelange-macherel, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with GFP fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles, and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting Peptide sequences and is suitable for animal and plant organelle-targeted proteins.
-
Experimental determination of organelle targeting-Peptide Cleavage sites using transient expression of green fluorescent protein translational fusions.
Analytical biochemistry, 2012Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Marie-hélène Avelange-macherel, Hélène Rogniaux, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence, which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with green fluorescent protein (GFP) fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting-Peptide sequences and is suitable for animal and plant organelle-targeted proteins.
-
Experimental determination of organelle targeting Peptide Cleavage sites using transient expression of GFP translational fusions
2012Co-Authors: Adrien Candat, Pauline Poupart, Jean-pierre Andrieu, Arnaud Chevrollier, Pascal Reynier, Helene Rogniaux-bonaventure, Marie-hélène Avelange-macherel, David MacherelAbstract:The majority of nuclear-encoded organellar proteins contain a cleavable presequence, which is necessary for protein targeting and import into the correct cellular compartment. Knowledge about targeting-Peptide Cleavage sites is essential for the structural and functional characterization of the mature organellar proteins as well as for a deeper understanding of the import process. Because of the low consensus and high variability of presequences, bioinformatics of targeting-Peptide Cleavage fails to predict the length of the targeting Peptide with high confidence. Therefore, we have developed a rapid and robust method to experimentally determine the Cleavage site of the transit Peptide for proteins imported into mitochondria or plastids. The protein precursor with green fluorescent protein (GFP) fused to its C-terminus is transiently expressed in cells (for animal proteins) or protoplasts (for plant proteins), allowing translocation into organelles and removal of the transit Peptide. After lysis, the matured protein is immunopurified using an anti-GFP antibody coupled to magnetic beads. The N-terminal amino sequence is then determined by Edman microsequencing or mass spectrometry. The method has been validated using proteins with known targeting-Peptide sequences and is suitable for animal and plant organelle-targeted proteins.