Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys

The transport of calcium to the extracellular space carried out by plasma membrane Ca2+ pumps (PMCAs) is essential for maintaining low Ca 2+ concentrations in the cytosol of eukaryotic cells. The activity of PMC Asis controlled by autoinhibition. Autoinhibition is relieved by the binding of Ca2+-cal...

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Autores principales:	Mazzitelli, L.R., Adamo, H.P.
Formato:	JOUR
Materias:	Calmodulin Cell membranes Molecules Pumps Autoinhibitory domain C-terminal segments Eukaryotic cells Extracellular space Intramolecular interactions Maximal velocity Negatively charged Transmembrane helices Calcium adenosine triphosphatase (calcium) calcium calmodulin glutamic acid green fluorescent protein h4xb plasma membrane calcium pump lysine unclassified drug amino acid substitution article controlled study cytosol enzyme activity enzyme conformation mutation priority journal ATPases Calcium ATPase Calcium Transport Enzymes Transporters Catalysis Cell Membrane Cytosol Gene Deletion Glutamine Green Fluorescent Proteins Humans Lysine Mutation Plasma Membrane Calcium-Transporting ATPases Protein Binding Protein Isoforms Protein Structure, Tertiary Saccharomyces cerevisiae
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00219258_v289_n15_p10761_Mazzitelli
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_00219258_v289_n15_p10761_Mazzitelli
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spelling	todo:paper_00219258_v289_n15_p10761_Mazzitelli2023-10-03T14:23:21Z Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys Mazzitelli, L.R. Adamo, H.P. Calmodulin Cell membranes Molecules Pumps Autoinhibitory domain C-terminal segments Eukaryotic cells Extracellular space Intramolecular interactions Maximal velocity Negatively charged Transmembrane helices Calcium adenosine triphosphatase (calcium) calcium calmodulin glutamic acid green fluorescent protein h4xb plasma membrane calcium pump lysine unclassified drug amino acid substitution article controlled study cytosol enzyme activity enzyme conformation mutation priority journal ATPases Calcium ATPase Calcium Transport Enzymes Transporters Calcium Calmodulin Catalysis Cell Membrane Cytosol Gene Deletion Glutamine Green Fluorescent Proteins Humans Lysine Mutation Plasma Membrane Calcium-Transporting ATPases Protein Binding Protein Isoforms Protein Structure, Tertiary Saccharomyces cerevisiae The transport of calcium to the extracellular space carried out by plasma membrane Ca2+ pumps (PMCAs) is essential for maintaining low Ca 2+ concentrations in the cytosol of eukaryotic cells. The activity of PMC Asis controlled by autoinhibition. Autoinhibition is relieved by the binding of Ca2+-calmodulin to the calmodulin-binding autoinhibitory sequence, which in the human PMCA is located in the C-terminal segment and results in a PMCA of high maximal velocity of transport and high affinity for Ca2+. Autoinhibition involves the intramolecular interaction between the autoinhibitory domain and a not well defined region of the molecule near the catalytic site. Here we show that the fusion of GFPto theCterminus of the h4xbPMCA causes partial loss of autoinhibition by specifically increasing the Vmax. Mutation of residue Glu99 to Lys in the cytosolic portion of the M1 transmembrane helix at the other end of the molecule brought the Vmax of the h4xbPMCAto near that of the calmodulin-activated enzyme without increasing the apparent affinity for Ca2+. Altogether, the results suggest that the autoinhibitory interaction of the extreme C-terminal segment of the h4PMCAis disturbed by changes of negatively charged residues of the N-terminal region. This would be consistent with a recently proposed model of an autoinhibited form of the plant ACA8 pump, although some differences are noted. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00219258_v289_n15_p10761_Mazzitelli
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Calmodulin Cell membranes Molecules Pumps Autoinhibitory domain C-terminal segments Eukaryotic cells Extracellular space Intramolecular interactions Maximal velocity Negatively charged Transmembrane helices Calcium adenosine triphosphatase (calcium) calcium calmodulin glutamic acid green fluorescent protein h4xb plasma membrane calcium pump lysine unclassified drug amino acid substitution article controlled study cytosol enzyme activity enzyme conformation mutation priority journal ATPases Calcium ATPase Calcium Transport Enzymes Transporters Calcium Calmodulin Catalysis Cell Membrane Cytosol Gene Deletion Glutamine Green Fluorescent Proteins Humans Lysine Mutation Plasma Membrane Calcium-Transporting ATPases Protein Binding Protein Isoforms Protein Structure, Tertiary Saccharomyces cerevisiae
spellingShingle	Calmodulin Cell membranes Molecules Pumps Autoinhibitory domain C-terminal segments Eukaryotic cells Extracellular space Intramolecular interactions Maximal velocity Negatively charged Transmembrane helices Calcium adenosine triphosphatase (calcium) calcium calmodulin glutamic acid green fluorescent protein h4xb plasma membrane calcium pump lysine unclassified drug amino acid substitution article controlled study cytosol enzyme activity enzyme conformation mutation priority journal ATPases Calcium ATPase Calcium Transport Enzymes Transporters Calcium Calmodulin Catalysis Cell Membrane Cytosol Gene Deletion Glutamine Green Fluorescent Proteins Humans Lysine Mutation Plasma Membrane Calcium-Transporting ATPases Protein Binding Protein Isoforms Protein Structure, Tertiary Saccharomyces cerevisiae Mazzitelli, L.R. Adamo, H.P. Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys
topic_facet	Calmodulin Cell membranes Molecules Pumps Autoinhibitory domain C-terminal segments Eukaryotic cells Extracellular space Intramolecular interactions Maximal velocity Negatively charged Transmembrane helices Calcium adenosine triphosphatase (calcium) calcium calmodulin glutamic acid green fluorescent protein h4xb plasma membrane calcium pump lysine unclassified drug amino acid substitution article controlled study cytosol enzyme activity enzyme conformation mutation priority journal ATPases Calcium ATPase Calcium Transport Enzymes Transporters Calcium Calmodulin Catalysis Cell Membrane Cytosol Gene Deletion Glutamine Green Fluorescent Proteins Humans Lysine Mutation Plasma Membrane Calcium-Transporting ATPases Protein Binding Protein Isoforms Protein Structure, Tertiary Saccharomyces cerevisiae
description	The transport of calcium to the extracellular space carried out by plasma membrane Ca2+ pumps (PMCAs) is essential for maintaining low Ca 2+ concentrations in the cytosol of eukaryotic cells. The activity of PMC Asis controlled by autoinhibition. Autoinhibition is relieved by the binding of Ca2+-calmodulin to the calmodulin-binding autoinhibitory sequence, which in the human PMCA is located in the C-terminal segment and results in a PMCA of high maximal velocity of transport and high affinity for Ca2+. Autoinhibition involves the intramolecular interaction between the autoinhibitory domain and a not well defined region of the molecule near the catalytic site. Here we show that the fusion of GFPto theCterminus of the h4xbPMCA causes partial loss of autoinhibition by specifically increasing the Vmax. Mutation of residue Glu99 to Lys in the cytosolic portion of the M1 transmembrane helix at the other end of the molecule brought the Vmax of the h4xbPMCAto near that of the calmodulin-activated enzyme without increasing the apparent affinity for Ca2+. Altogether, the results suggest that the autoinhibitory interaction of the extreme C-terminal segment of the h4PMCAis disturbed by changes of negatively charged residues of the N-terminal region. This would be consistent with a recently proposed model of an autoinhibited form of the plant ACA8 pump, although some differences are noted. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.
format	JOUR
author	Mazzitelli, L.R. Adamo, H.P.
author_facet	Mazzitelli, L.R. Adamo, H.P.
author_sort	Mazzitelli, L.R.
title	Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys
title_short	Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys
title_full	Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys
title_fullStr	Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys
title_full_unstemmed	Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys
title_sort	hyperactivation of the human plasma membrane ca2+ pump pmca h4xb by mutation of glu99 to lys
url	http://hdl.handle.net/20.500.12110/paper_00219258_v289_n15_p10761_Mazzitelli
work_keys_str_mv	AT mazzitellilr hyperactivationofthehumanplasmamembraneca2pumppmcah4xbbymutationofglu99tolys AT adamohp hyperactivationofthehumanplasmamembraneca2pumppmcah4xbbymutationofglu99tolys
_version_	1807320896493322240

Hyperactivation of the human plasma membrane Ca2+ pump PMCA h4xb by mutation of Glu99 to Lys

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