The Experts below are selected from a list of 858 Experts worldwide ranked by ideXlab platform
Taro Kihara - One of the best experts on this subject based on the ideXlab platform.
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partial purification and characterization of sphingosine n acyltransferase ceramide synthase from bovine Liver Mitochondrion rich fraction
Lipids, 1998Co-Authors: Hiroshi Shimeno, Shinji Soeda, Masahiko Sakamoto, Takeshi Kouchi, Takashi Kowakame, Taro KiharaAbstract:Sphingosine N-acyltransferase (ceramide synthase, E.C. 2.3.1.24) was solubilized from bovine Liver Mitochondrion-rich fraction with n-ocytl β-d-thioglucoside as the detergent and partially purified by sequential chromatography on columns of DE-32, shingosine affinity, and Sepharose CL-6B. The partially purified preparation migrated on SDS-polyacrylamide gel electrophoresis as two major protein bands of 62 and 72 kDa. The molecular mass of the enzyme estimated by gel filtration was 240–260 kDa, suggesting that the partially purified enzyme is present in a subunit form or simply has an aggregative nature. The specific activity of the final preparation for the condensation of sphingosine with stearoyl-CoA increased by 98.7-fold compared with the starting material. The optimal pH value for the ceramide synthesis was 7.5. The partially purified enzyme had an apparent K m of 146 μM and a V max of 11.1 nmol/min/mg protein for stearoyl-CoA. The K m and V max values toward sphingosine were 171 μM and 11.3 nmol/min/mg protein, respectively. Interestingly, sphinganine was also a good substrate for this enzyme, and the K m and V max values were 144 μM and 8.5 nmol/min/mg protein, respectively.
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partial purification and characterization of sphingosine n acyltransferase ceramide synthase from bovine Liver Mitochondrion rich fraction
Lipids, 1998Co-Authors: Hiroshi Shimeno, Shinji Soeda, Masahiko Sakamoto, Takeshi Kouchi, Takashi Kowakame, Taro KiharaAbstract:Sphingosine N-acyltransferase (ceramide synthase, E.C. 2.3.1.24) was solubilized from bovine Liver Mitochondrion-rich fraction with n-octyl beta-D-thioglucoside as the detergent and partially purified by sequential chromatography on columns of DE-32, shingosine affinity, and Sepharose CL-6B. The partially purified preparation migrated on SDS-polyacrylamide gel electrophoresis as two major protein bands of 62 and 72 kDa. The molecular mass of the enzyme estimated by gel filtration was 240-260 kDa, suggesting that the partially purified enzyme is present in a subunit form or simply has an aggregative nature. The specific activity of the final preparation for the condensation of sphingosine with stearoyl-CoA increased by 98.7-fold compared with the starting material. The optimal pH value for the ceramide synthesis was 7.5. The partially purified enzyme had an apparent Km of 146 microM and a Vmax of 11.1 nmol/min/mg protein for stearoyl-CoA. The Km and Vmax values toward sphingosine were 171 microM and 11.3 nmol/min/mg protein, respectively. Interestingly, sphinganine was also a good substrate for this enzyme, and the Km and Vmax values were 144 microM and 8.5 nmol/min/mg protein, respectively.
Hofestädt Ralf - One of the best experts on this subject based on the ideXlab platform.
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Prediction of thioredoxin and glutaredoxin target proteins by identifying reversibly oxidized cysteinyl residues
De Gruyter, 2010Co-Authors: Lee Hang-mao, Dietz Karl Josef, Hofestädt RalfAbstract:A significant part of cellular proteins undergo reversible thiol-dependent redox transitions which often control or switch protein functions. Thioredoxins and glutaredoxins constitute two key players in this redox regulatory protein network. Both interact with various categories of proteins containing reversibly oxidized cysteinyl residues. The identification of thioredoxin/glutaredoxin target proteins is a critical step in constructing the redox regulatory network of cells or subcellular compartments. Due to the scarcity of thioredoxin/glutaredoxin target protein records in the public database, a tool called Reversibly Oxidized Cysteine Detector (ROCD) is implemented here to identify potential thioredoxin/glutaredoxin target proteins computationally, so that the in silico construction of redox regulatory network may become feasible. ROCD was tested on 46 thioredoxin target proteins in plant Mitochondrion, and the recall rate was 66.7% when 50% sequence identity was chosen for structural model selection. ROCD will be used to predict the thioredoxin/glutaredoxin target proteins in human Liver Mitochondrion for our redox regulatory network construction project. The ROCD will be developed further to provide prediction with more reliability and incorporated into biological network visualization tools as a node prediction component. This work will advance the capability of traditional database- or text mining-based method in the network construction
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Prediction of thioredoxin and glutaredoxin target proteins by identifying reversibly oxidized cysteinyl residues.
IMBio Informationsmanagement in der Biotechnologie e.V., 2010Co-Authors: Lee Hang-mao, Dietz Karl-josef, Hofestädt RalfAbstract:Lee H-M, Dietz K-J, Hofestädt R. Prediction of thioredoxin and glutaredoxin target proteins by identifying reversibly oxidized cysteinyl residues. Journal of Integrative Bioinformatics (Special Issue). 2010;7(3):23-34.A significant part of cellular proteins undergo reversible thiol-dependent redox transitions which often control or switch protein functions. Thioredoxins and glutaredoxins constitute two key players in this redox regulatory protein network. Both interact with various categories of proteins containing reversibly oxidized cysteinyl residues. The identification of thioredoxin/glutaredoxin target proteins is a critical step in constructing the redox regulatory network of cells or subcellular compartments. Due to the scarcity of thioredoxin/glutaredoxin target protein records in the public database, a tool called Reversibly Oxidized Cysteine Detector (ROCD) is implemented here to identify potential thioredoxin/glutaredoxin target proteins computationally, so that the in silico construction of redox regulatory network may become feasible. ROCD was tested on 46 thioredoxin target proteins in plant Mitochondrion, and the recall rate was 66.7% when 50% sequence identity was chosen for structural model selection. ROCD will be used to predict the thioredoxin/glutaredoxin target proteins in human Liver Mitochondrion for our redox regulatory network construction project. The ROCD will be developed further to provide prediction with more reliability and incorporated into biological network visualization tools as a node prediction component. This work will advance the capability of traditional database- or text mining-based method in the network construction
Hiroshi Shimeno - One of the best experts on this subject based on the ideXlab platform.
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partial purification and characterization of sphingosine n acyltransferase ceramide synthase from bovine Liver Mitochondrion rich fraction
Lipids, 1998Co-Authors: Hiroshi Shimeno, Shinji Soeda, Masahiko Sakamoto, Takeshi Kouchi, Takashi Kowakame, Taro KiharaAbstract:Sphingosine N-acyltransferase (ceramide synthase, E.C. 2.3.1.24) was solubilized from bovine Liver Mitochondrion-rich fraction with n-ocytl β-d-thioglucoside as the detergent and partially purified by sequential chromatography on columns of DE-32, shingosine affinity, and Sepharose CL-6B. The partially purified preparation migrated on SDS-polyacrylamide gel electrophoresis as two major protein bands of 62 and 72 kDa. The molecular mass of the enzyme estimated by gel filtration was 240–260 kDa, suggesting that the partially purified enzyme is present in a subunit form or simply has an aggregative nature. The specific activity of the final preparation for the condensation of sphingosine with stearoyl-CoA increased by 98.7-fold compared with the starting material. The optimal pH value for the ceramide synthesis was 7.5. The partially purified enzyme had an apparent K m of 146 μM and a V max of 11.1 nmol/min/mg protein for stearoyl-CoA. The K m and V max values toward sphingosine were 171 μM and 11.3 nmol/min/mg protein, respectively. Interestingly, sphinganine was also a good substrate for this enzyme, and the K m and V max values were 144 μM and 8.5 nmol/min/mg protein, respectively.
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partial purification and characterization of sphingosine n acyltransferase ceramide synthase from bovine Liver Mitochondrion rich fraction
Lipids, 1998Co-Authors: Hiroshi Shimeno, Shinji Soeda, Masahiko Sakamoto, Takeshi Kouchi, Takashi Kowakame, Taro KiharaAbstract:Sphingosine N-acyltransferase (ceramide synthase, E.C. 2.3.1.24) was solubilized from bovine Liver Mitochondrion-rich fraction with n-octyl beta-D-thioglucoside as the detergent and partially purified by sequential chromatography on columns of DE-32, shingosine affinity, and Sepharose CL-6B. The partially purified preparation migrated on SDS-polyacrylamide gel electrophoresis as two major protein bands of 62 and 72 kDa. The molecular mass of the enzyme estimated by gel filtration was 240-260 kDa, suggesting that the partially purified enzyme is present in a subunit form or simply has an aggregative nature. The specific activity of the final preparation for the condensation of sphingosine with stearoyl-CoA increased by 98.7-fold compared with the starting material. The optimal pH value for the ceramide synthesis was 7.5. The partially purified enzyme had an apparent Km of 146 microM and a Vmax of 11.1 nmol/min/mg protein for stearoyl-CoA. The Km and Vmax values toward sphingosine were 171 microM and 11.3 nmol/min/mg protein, respectively. Interestingly, sphinganine was also a good substrate for this enzyme, and the Km and Vmax values were 144 microM and 8.5 nmol/min/mg protein, respectively.
Lee Hang-mao - One of the best experts on this subject based on the ideXlab platform.
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Prediction of thioredoxin and glutaredoxin target proteins by identifying reversibly oxidized cysteinyl residues
De Gruyter, 2010Co-Authors: Lee Hang-mao, Dietz Karl Josef, Hofestädt RalfAbstract:A significant part of cellular proteins undergo reversible thiol-dependent redox transitions which often control or switch protein functions. Thioredoxins and glutaredoxins constitute two key players in this redox regulatory protein network. Both interact with various categories of proteins containing reversibly oxidized cysteinyl residues. The identification of thioredoxin/glutaredoxin target proteins is a critical step in constructing the redox regulatory network of cells or subcellular compartments. Due to the scarcity of thioredoxin/glutaredoxin target protein records in the public database, a tool called Reversibly Oxidized Cysteine Detector (ROCD) is implemented here to identify potential thioredoxin/glutaredoxin target proteins computationally, so that the in silico construction of redox regulatory network may become feasible. ROCD was tested on 46 thioredoxin target proteins in plant Mitochondrion, and the recall rate was 66.7% when 50% sequence identity was chosen for structural model selection. ROCD will be used to predict the thioredoxin/glutaredoxin target proteins in human Liver Mitochondrion for our redox regulatory network construction project. The ROCD will be developed further to provide prediction with more reliability and incorporated into biological network visualization tools as a node prediction component. This work will advance the capability of traditional database- or text mining-based method in the network construction
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Prediction of thioredoxin and glutaredoxin target proteins by identifying reversibly oxidized cysteinyl residues.
IMBio Informationsmanagement in der Biotechnologie e.V., 2010Co-Authors: Lee Hang-mao, Dietz Karl-josef, Hofestädt RalfAbstract:Lee H-M, Dietz K-J, Hofestädt R. Prediction of thioredoxin and glutaredoxin target proteins by identifying reversibly oxidized cysteinyl residues. Journal of Integrative Bioinformatics (Special Issue). 2010;7(3):23-34.A significant part of cellular proteins undergo reversible thiol-dependent redox transitions which often control or switch protein functions. Thioredoxins and glutaredoxins constitute two key players in this redox regulatory protein network. Both interact with various categories of proteins containing reversibly oxidized cysteinyl residues. The identification of thioredoxin/glutaredoxin target proteins is a critical step in constructing the redox regulatory network of cells or subcellular compartments. Due to the scarcity of thioredoxin/glutaredoxin target protein records in the public database, a tool called Reversibly Oxidized Cysteine Detector (ROCD) is implemented here to identify potential thioredoxin/glutaredoxin target proteins computationally, so that the in silico construction of redox regulatory network may become feasible. ROCD was tested on 46 thioredoxin target proteins in plant Mitochondrion, and the recall rate was 66.7% when 50% sequence identity was chosen for structural model selection. ROCD will be used to predict the thioredoxin/glutaredoxin target proteins in human Liver Mitochondrion for our redox regulatory network construction project. The ROCD will be developed further to provide prediction with more reliability and incorporated into biological network visualization tools as a node prediction component. This work will advance the capability of traditional database- or text mining-based method in the network construction
Shinji Soeda - One of the best experts on this subject based on the ideXlab platform.
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partial purification and characterization of sphingosine n acyltransferase ceramide synthase from bovine Liver Mitochondrion rich fraction
Lipids, 1998Co-Authors: Hiroshi Shimeno, Shinji Soeda, Masahiko Sakamoto, Takeshi Kouchi, Takashi Kowakame, Taro KiharaAbstract:Sphingosine N-acyltransferase (ceramide synthase, E.C. 2.3.1.24) was solubilized from bovine Liver Mitochondrion-rich fraction with n-ocytl β-d-thioglucoside as the detergent and partially purified by sequential chromatography on columns of DE-32, shingosine affinity, and Sepharose CL-6B. The partially purified preparation migrated on SDS-polyacrylamide gel electrophoresis as two major protein bands of 62 and 72 kDa. The molecular mass of the enzyme estimated by gel filtration was 240–260 kDa, suggesting that the partially purified enzyme is present in a subunit form or simply has an aggregative nature. The specific activity of the final preparation for the condensation of sphingosine with stearoyl-CoA increased by 98.7-fold compared with the starting material. The optimal pH value for the ceramide synthesis was 7.5. The partially purified enzyme had an apparent K m of 146 μM and a V max of 11.1 nmol/min/mg protein for stearoyl-CoA. The K m and V max values toward sphingosine were 171 μM and 11.3 nmol/min/mg protein, respectively. Interestingly, sphinganine was also a good substrate for this enzyme, and the K m and V max values were 144 μM and 8.5 nmol/min/mg protein, respectively.
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partial purification and characterization of sphingosine n acyltransferase ceramide synthase from bovine Liver Mitochondrion rich fraction
Lipids, 1998Co-Authors: Hiroshi Shimeno, Shinji Soeda, Masahiko Sakamoto, Takeshi Kouchi, Takashi Kowakame, Taro KiharaAbstract:Sphingosine N-acyltransferase (ceramide synthase, E.C. 2.3.1.24) was solubilized from bovine Liver Mitochondrion-rich fraction with n-octyl beta-D-thioglucoside as the detergent and partially purified by sequential chromatography on columns of DE-32, shingosine affinity, and Sepharose CL-6B. The partially purified preparation migrated on SDS-polyacrylamide gel electrophoresis as two major protein bands of 62 and 72 kDa. The molecular mass of the enzyme estimated by gel filtration was 240-260 kDa, suggesting that the partially purified enzyme is present in a subunit form or simply has an aggregative nature. The specific activity of the final preparation for the condensation of sphingosine with stearoyl-CoA increased by 98.7-fold compared with the starting material. The optimal pH value for the ceramide synthesis was 7.5. The partially purified enzyme had an apparent Km of 146 microM and a Vmax of 11.1 nmol/min/mg protein for stearoyl-CoA. The Km and Vmax values toward sphingosine were 171 microM and 11.3 nmol/min/mg protein, respectively. Interestingly, sphinganine was also a good substrate for this enzyme, and the Km and Vmax values were 144 microM and 8.5 nmol/min/mg protein, respectively.