Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity

The TPM domain constitutes a family of recently characterized protein domains that are present in most living organisms. Although some progress has been made in understanding the cellular role of TPM-containing proteins, the relationship between structure and function is not clear yet. We have recen...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Smal, Clara, Ithuralde, Raul Esteban, Turjanski, Adrián Gustavo, Cicero, Daniel Oscar, Arán, Martín
Publicado:	2016
Materias:	Antarctic bacteria Bizionia argentinensis nuclear magnetic resonance phosphatase activity structural genomics TPM domain calcium ion magnesium ion phosphatase bacterial protein calcium divalent cation magnesium metal protein binding Article carboxy terminal sequence conformational transition enzyme active site enzyme activity enzyme binding enzyme mechanism heteronuclear single quantum coherence limit of quantitation low temperature molecular dynamics nitrogen nuclear magnetic resonance nonhuman priority journal protein unfolding adaptation amino acid sequence Antarctica binding site chemistry cold enzyme stability enzymology Flavobacteriaceae genetics kinetics metabolism molecular model mutation nuclear magnetic resonance spectroscopy pH protein domain sequence homology structure activity relation Adaptation, Physiological Amino Acid Sequence Antarctic Regions Bacterial Proteins Binding Sites Calcium Cations, Divalent Cold Temperature Enzyme Stability Hydrogen-Ion Concentration Kinetics Magnesium Magnetic Resonance Spectroscopy Metals Models, Molecular Mutation Phosphoric Monoester Hydrolases Protein Binding Protein Domains Sequence Homology, Amino Acid Structure-Activity Relationship
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_1742464X_v283_n23_p4370_Pellizza http://hdl.handle.net/20.500.12110/paper_1742464X_v283_n23_p4370_Pellizza
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_1742464X_v283_n23_p4370_Pellizza
record_format	dspace
spelling	paper:paper_1742464X_v283_n23_p4370_Pellizza2023-06-08T16:27:04Z Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity Smal, Clara Ithuralde, Raul Esteban Turjanski, Adrián Gustavo Cicero, Daniel Oscar Arán, Martín Antarctic bacteria Bizionia argentinensis nuclear magnetic resonance phosphatase activity structural genomics TPM domain calcium ion magnesium ion phosphatase bacterial protein calcium divalent cation magnesium metal phosphatase protein binding Article carboxy terminal sequence conformational transition enzyme active site enzyme activity enzyme binding enzyme mechanism heteronuclear single quantum coherence limit of quantitation low temperature molecular dynamics nitrogen nuclear magnetic resonance nonhuman priority journal protein unfolding adaptation amino acid sequence Antarctica binding site chemistry cold enzyme stability enzymology Flavobacteriaceae genetics kinetics metabolism molecular model mutation nuclear magnetic resonance spectroscopy pH protein domain sequence homology structure activity relation Adaptation, Physiological Amino Acid Sequence Antarctic Regions Bacterial Proteins Binding Sites Calcium Cations, Divalent Cold Temperature Enzyme Stability Flavobacteriaceae Hydrogen-Ion Concentration Kinetics Magnesium Magnetic Resonance Spectroscopy Metals Models, Molecular Mutation Phosphoric Monoester Hydrolases Protein Binding Protein Domains Sequence Homology, Amino Acid Structure-Activity Relationship The TPM domain constitutes a family of recently characterized protein domains that are present in most living organisms. Although some progress has been made in understanding the cellular role of TPM-containing proteins, the relationship between structure and function is not clear yet. We have recently solved the solution and crystal structure of one TPM domain (BA42) from the Antarctic bacterium Bizionia argentinensis. In this work, we demonstrate that BA42 has phosphoric-monoester hydrolase activity. The activity of BA42 is strictly dependent on the binding of divalent metals and retains nearly 70% of the maximum at 4 °C, a typical characteristic of cold-adapted enzymes. From HSQC, 15N relaxation measurements, and molecular dynamics studies, we determine that the flexibility of the crossing loops was associated to the protein activity. Thermal unfolding experiments showed that the local increment in flexibility of Mg2+-bound BA42, when compared with Ca2+-bound BA42, is associated to a decrease in global protein stability. Finally, through mutagenesis experiments, we unambiguously demonstrate that the region comprising the metal-binding site participates in the catalytic mechanism. The results shown here contribute to the understanding of the relationship between structure and function of this new family of TPM domains providing important cues on the regulatory role of Mg2+ and Ca2+ and the molecular mechanism underlying enzyme activity at low temperatures. © 2016 Federation of European Biochemical Societies Fil:Smal, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ithuralde, R.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Turjanski, A.G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Cicero, D.O. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Arán, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2016 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_1742464X_v283_n23_p4370_Pellizza http://hdl.handle.net/20.500.12110/paper_1742464X_v283_n23_p4370_Pellizza
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Antarctic bacteria Bizionia argentinensis nuclear magnetic resonance phosphatase activity structural genomics TPM domain calcium ion magnesium ion phosphatase bacterial protein calcium divalent cation magnesium metal phosphatase protein binding Article carboxy terminal sequence conformational transition enzyme active site enzyme activity enzyme binding enzyme mechanism heteronuclear single quantum coherence limit of quantitation low temperature molecular dynamics nitrogen nuclear magnetic resonance nonhuman priority journal protein unfolding adaptation amino acid sequence Antarctica binding site chemistry cold enzyme stability enzymology Flavobacteriaceae genetics kinetics metabolism molecular model mutation nuclear magnetic resonance spectroscopy pH protein domain sequence homology structure activity relation Adaptation, Physiological Amino Acid Sequence Antarctic Regions Bacterial Proteins Binding Sites Calcium Cations, Divalent Cold Temperature Enzyme Stability Flavobacteriaceae Hydrogen-Ion Concentration Kinetics Magnesium Magnetic Resonance Spectroscopy Metals Models, Molecular Mutation Phosphoric Monoester Hydrolases Protein Binding Protein Domains Sequence Homology, Amino Acid Structure-Activity Relationship
spellingShingle	Antarctic bacteria Bizionia argentinensis nuclear magnetic resonance phosphatase activity structural genomics TPM domain calcium ion magnesium ion phosphatase bacterial protein calcium divalent cation magnesium metal phosphatase protein binding Article carboxy terminal sequence conformational transition enzyme active site enzyme activity enzyme binding enzyme mechanism heteronuclear single quantum coherence limit of quantitation low temperature molecular dynamics nitrogen nuclear magnetic resonance nonhuman priority journal protein unfolding adaptation amino acid sequence Antarctica binding site chemistry cold enzyme stability enzymology Flavobacteriaceae genetics kinetics metabolism molecular model mutation nuclear magnetic resonance spectroscopy pH protein domain sequence homology structure activity relation Adaptation, Physiological Amino Acid Sequence Antarctic Regions Bacterial Proteins Binding Sites Calcium Cations, Divalent Cold Temperature Enzyme Stability Flavobacteriaceae Hydrogen-Ion Concentration Kinetics Magnesium Magnetic Resonance Spectroscopy Metals Models, Molecular Mutation Phosphoric Monoester Hydrolases Protein Binding Protein Domains Sequence Homology, Amino Acid Structure-Activity Relationship Smal, Clara Ithuralde, Raul Esteban Turjanski, Adrián Gustavo Cicero, Daniel Oscar Arán, Martín Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity
topic_facet	Antarctic bacteria Bizionia argentinensis nuclear magnetic resonance phosphatase activity structural genomics TPM domain calcium ion magnesium ion phosphatase bacterial protein calcium divalent cation magnesium metal phosphatase protein binding Article carboxy terminal sequence conformational transition enzyme active site enzyme activity enzyme binding enzyme mechanism heteronuclear single quantum coherence limit of quantitation low temperature molecular dynamics nitrogen nuclear magnetic resonance nonhuman priority journal protein unfolding adaptation amino acid sequence Antarctica binding site chemistry cold enzyme stability enzymology Flavobacteriaceae genetics kinetics metabolism molecular model mutation nuclear magnetic resonance spectroscopy pH protein domain sequence homology structure activity relation Adaptation, Physiological Amino Acid Sequence Antarctic Regions Bacterial Proteins Binding Sites Calcium Cations, Divalent Cold Temperature Enzyme Stability Flavobacteriaceae Hydrogen-Ion Concentration Kinetics Magnesium Magnetic Resonance Spectroscopy Metals Models, Molecular Mutation Phosphoric Monoester Hydrolases Protein Binding Protein Domains Sequence Homology, Amino Acid Structure-Activity Relationship
description	The TPM domain constitutes a family of recently characterized protein domains that are present in most living organisms. Although some progress has been made in understanding the cellular role of TPM-containing proteins, the relationship between structure and function is not clear yet. We have recently solved the solution and crystal structure of one TPM domain (BA42) from the Antarctic bacterium Bizionia argentinensis. In this work, we demonstrate that BA42 has phosphoric-monoester hydrolase activity. The activity of BA42 is strictly dependent on the binding of divalent metals and retains nearly 70% of the maximum at 4 °C, a typical characteristic of cold-adapted enzymes. From HSQC, 15N relaxation measurements, and molecular dynamics studies, we determine that the flexibility of the crossing loops was associated to the protein activity. Thermal unfolding experiments showed that the local increment in flexibility of Mg2+-bound BA42, when compared with Ca2+-bound BA42, is associated to a decrease in global protein stability. Finally, through mutagenesis experiments, we unambiguously demonstrate that the region comprising the metal-binding site participates in the catalytic mechanism. The results shown here contribute to the understanding of the relationship between structure and function of this new family of TPM domains providing important cues on the regulatory role of Mg2+ and Ca2+ and the molecular mechanism underlying enzyme activity at low temperatures. © 2016 Federation of European Biochemical Societies
author	Smal, Clara Ithuralde, Raul Esteban Turjanski, Adrián Gustavo Cicero, Daniel Oscar Arán, Martín
author_facet	Smal, Clara Ithuralde, Raul Esteban Turjanski, Adrián Gustavo Cicero, Daniel Oscar Arán, Martín
author_sort	Smal, Clara
title	Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity
title_short	Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity
title_full	Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity
title_fullStr	Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity
title_full_unstemmed	Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity
title_sort	structural and functional characterization of a cold-adapted stand-alone tpm domain reveals a relationship between dynamics and phosphatase activity
publishDate	2016
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_1742464X_v283_n23_p4370_Pellizza http://hdl.handle.net/20.500.12110/paper_1742464X_v283_n23_p4370_Pellizza
work_keys_str_mv	AT smalclara structuralandfunctionalcharacterizationofacoldadaptedstandalonetpmdomainrevealsarelationshipbetweendynamicsandphosphataseactivity AT ithuralderaulesteban structuralandfunctionalcharacterizationofacoldadaptedstandalonetpmdomainrevealsarelationshipbetweendynamicsandphosphataseactivity AT turjanskiadriangustavo structuralandfunctionalcharacterizationofacoldadaptedstandalonetpmdomainrevealsarelationshipbetweendynamicsandphosphataseactivity AT cicerodanieloscar structuralandfunctionalcharacterizationofacoldadaptedstandalonetpmdomainrevealsarelationshipbetweendynamicsandphosphataseactivity AT aranmartin structuralandfunctionalcharacterizationofacoldadaptedstandalonetpmdomainrevealsarelationshipbetweendynamicsandphosphataseactivity
_version_	1768546557331767296

Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity

Ejemplares similares