Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase

A system was developed for testing the effect of phospholipids on dolichyl phosphate phosphatase, a membrane-associated enzyme. This enzyme was solubilized, delipidated, stabilized and concentrated in such a way that minimal quantities of Triton X-100 were carried by enzyme extracts to the incubatio...

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Autores principales:	Boscoboinik, Daniel Osvaldo, Belocopitow, Enrique
Publicado:	1984
Materias:	(Rat liver microsome) Detergent Dolichyl phosphate phosphatase Enzyme assay Phospholipid phospholipid animal cell dolichyl phosphatase enzyme assay liver microsome nonhuman rat Animal Catalase Cytochrome c Detergents Intracellular Membranes Kinetics Microsomes, Liver Octoxynol Phospholipids Phosphoric Monoester Hydrolases Polyethylene Glycols Rats Support, Non-U.S. Gov't Surface-Active Agents
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00052760_v794_n1_p41_Boscoboinik http://hdl.handle.net/20.500.12110/paper_00052760_v794_n1_p41_Boscoboinik
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_00052760_v794_n1_p41_Boscoboinik
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spelling	paper:paper_00052760_v794_n1_p41_Boscoboinik2023-06-08T14:29:56Z Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase Boscoboinik, Daniel Osvaldo Belocopitow, Enrique (Rat liver microsome) Detergent Dolichyl phosphate phosphatase Enzyme assay Phospholipid phospholipid animal cell dolichyl phosphatase enzyme assay liver microsome nonhuman rat Animal Catalase Cytochrome c Detergents Intracellular Membranes Kinetics Microsomes, Liver Octoxynol Phospholipids Phosphoric Monoester Hydrolases Polyethylene Glycols Rats Support, Non-U.S. Gov't Surface-Active Agents A system was developed for testing the effect of phospholipids on dolichyl phosphate phosphatase, a membrane-associated enzyme. This enzyme was solubilized, delipidated, stabilized and concentrated in such a way that minimal quantities of Triton X-100 were carried by enzyme extracts to the incubation mixture. Its substrate, dolichyl phosphate, could be kept in aqueous medium as suspended particles without addition of detergent. When dolichyl phosphate phosphatase was assayed using the substrate in this detergent-free form, values for Km, pH optimum and temperature optimum were different from those obtained with detergentsolubilized substrate. This assay of dolichyl phosphate phosphatase almost free of detergent allowed testing of the effect of specific phospholipids on enzyme activity with minimal interference produced by endogenous phospholipids or exogenous detergent. Sphingomyelin, phosphatidylethanolamine or phosphatidylcholine (zwitterionic phospholipids) acted as activators, whereas phosphatidic acid and phosphatidylinositol, negatively-charged phospholipids, were inhibitors of dolichyl phosphate phosphatase. © 1984. Fil:Boscoboinik, D.O. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Belocopitow, E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1984 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00052760_v794_n1_p41_Boscoboinik http://hdl.handle.net/20.500.12110/paper_00052760_v794_n1_p41_Boscoboinik
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	(Rat liver microsome) Detergent Dolichyl phosphate phosphatase Enzyme assay Phospholipid phospholipid animal cell dolichyl phosphatase enzyme assay liver microsome nonhuman rat Animal Catalase Cytochrome c Detergents Intracellular Membranes Kinetics Microsomes, Liver Octoxynol Phospholipids Phosphoric Monoester Hydrolases Polyethylene Glycols Rats Support, Non-U.S. Gov't Surface-Active Agents
spellingShingle	(Rat liver microsome) Detergent Dolichyl phosphate phosphatase Enzyme assay Phospholipid phospholipid animal cell dolichyl phosphatase enzyme assay liver microsome nonhuman rat Animal Catalase Cytochrome c Detergents Intracellular Membranes Kinetics Microsomes, Liver Octoxynol Phospholipids Phosphoric Monoester Hydrolases Polyethylene Glycols Rats Support, Non-U.S. Gov't Surface-Active Agents Boscoboinik, Daniel Osvaldo Belocopitow, Enrique Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase
topic_facet	(Rat liver microsome) Detergent Dolichyl phosphate phosphatase Enzyme assay Phospholipid phospholipid animal cell dolichyl phosphatase enzyme assay liver microsome nonhuman rat Animal Catalase Cytochrome c Detergents Intracellular Membranes Kinetics Microsomes, Liver Octoxynol Phospholipids Phosphoric Monoester Hydrolases Polyethylene Glycols Rats Support, Non-U.S. Gov't Surface-Active Agents
description	A system was developed for testing the effect of phospholipids on dolichyl phosphate phosphatase, a membrane-associated enzyme. This enzyme was solubilized, delipidated, stabilized and concentrated in such a way that minimal quantities of Triton X-100 were carried by enzyme extracts to the incubation mixture. Its substrate, dolichyl phosphate, could be kept in aqueous medium as suspended particles without addition of detergent. When dolichyl phosphate phosphatase was assayed using the substrate in this detergent-free form, values for Km, pH optimum and temperature optimum were different from those obtained with detergentsolubilized substrate. This assay of dolichyl phosphate phosphatase almost free of detergent allowed testing of the effect of specific phospholipids on enzyme activity with minimal interference produced by endogenous phospholipids or exogenous detergent. Sphingomyelin, phosphatidylethanolamine or phosphatidylcholine (zwitterionic phospholipids) acted as activators, whereas phosphatidic acid and phosphatidylinositol, negatively-charged phospholipids, were inhibitors of dolichyl phosphate phosphatase. © 1984.
author	Boscoboinik, Daniel Osvaldo Belocopitow, Enrique
author_facet	Boscoboinik, Daniel Osvaldo Belocopitow, Enrique
author_sort	Boscoboinik, Daniel Osvaldo
title	Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase
title_short	Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase
title_full	Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase
title_fullStr	Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase
title_full_unstemmed	Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase
title_sort	dolichyl phosphate phosphatase in rat liver microsomes. avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase
publishDate	1984
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00052760_v794_n1_p41_Boscoboinik http://hdl.handle.net/20.500.12110/paper_00052760_v794_n1_p41_Boscoboinik
work_keys_str_mv	AT boscoboinikdanielosvaldo dolichylphosphatephosphataseinratlivermicrosomesavoidanceoftheuseofdetergentintestingtheeffectofphospholipidsondolichylphosphatephosphatase AT belocopitowenrique dolichylphosphatephosphataseinratlivermicrosomesavoidanceoftheuseofdetergentintestingtheeffectofphospholipidsondolichylphosphatephosphatase
_version_	1768541873889083392

Dolichyl phosphate phosphatase in rat liver microsomes. Avoidance of the use of detergent in testing the effect of phospholipids on dolichyl phosphate phosphatase

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