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...
Published in: | The FEBS Journal |
---|---|
Main Authors: | , , , , |
Other Authors: | , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Blackwell Publishing Ltd
2016
|
Subjects: | |
Online Access: | http://hdl.handle.net/2108/181089 https://doi.org/10.1111/febs.13929 |
id |
ftunivromatorver:oai:art.torvergata.it:2108/181089 |
---|---|
record_format |
openpolar |
spelling |
ftunivromatorver:oai:art.torvergata.it:2108/181089 2024-02-27T08:35:12+00:00 Structural and functional characterization of a cold-adapted stand-alone TPM domain reveals a relationship between dynamics and phosphatase activity Pellizza, L Smal, C Ithurralde, RE Turjanski, AG CICERO, DANIEL OSCAR Pellizza, L Smal, C Ithurralde, R Turjanski, A Cicero, Do 2016 http://hdl.handle.net/2108/181089 https://doi.org/10.1111/febs.13929 eng eng Blackwell Publishing Ltd info:eu-repo/semantics/altIdentifier/pmid/27754607 info:eu-repo/semantics/altIdentifier/wos/WOS:000392742500013 volume:283 issue:23 firstpage:4370 lastpage:4385 numberofpages:16 journal:THE FEBS JOURNAL http://hdl.handle.net/2108/181089 doi:10.1111/febs.13929 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85000542405 Antarctic bacteria Bizionia argentinensi nuclear magnetic resonance phosphatase activity structural genomic TPM domain Settore BIO/10 - BIOCHIMICA info:eu-repo/semantics/article 2016 ftunivromatorver https://doi.org/10.1111/febs.13929 2024-01-31T00:11:56Z 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. Article in Journal/Newspaper Antarc* Antarctic Universitá degli Studi di Roma "Tor Vergata": ART - Archivio Istituzionale della Ricerca Antarctic The Antarctic The FEBS Journal 283 23 4370 4385 |
institution |
Open Polar |
collection |
Universitá degli Studi di Roma "Tor Vergata": ART - Archivio Istituzionale della Ricerca |
op_collection_id |
ftunivromatorver |
language |
English |
topic |
Antarctic bacteria Bizionia argentinensi nuclear magnetic resonance phosphatase activity structural genomic TPM domain Settore BIO/10 - BIOCHIMICA |
spellingShingle |
Antarctic bacteria Bizionia argentinensi nuclear magnetic resonance phosphatase activity structural genomic TPM domain Settore BIO/10 - BIOCHIMICA Pellizza, L Smal, C Ithurralde, RE Turjanski, AG CICERO, DANIEL OSCAR 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 argentinensi nuclear magnetic resonance phosphatase activity structural genomic TPM domain Settore BIO/10 - BIOCHIMICA |
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. |
author2 |
Pellizza, L Smal, C Ithurralde, R Turjanski, A Cicero, Do |
format |
Article in Journal/Newspaper |
author |
Pellizza, L Smal, C Ithurralde, RE Turjanski, AG CICERO, DANIEL OSCAR |
author_facet |
Pellizza, L Smal, C Ithurralde, RE Turjanski, AG CICERO, DANIEL OSCAR |
author_sort |
Pellizza, L |
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 |
publisher |
Blackwell Publishing Ltd |
publishDate |
2016 |
url |
http://hdl.handle.net/2108/181089 https://doi.org/10.1111/febs.13929 |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
info:eu-repo/semantics/altIdentifier/pmid/27754607 info:eu-repo/semantics/altIdentifier/wos/WOS:000392742500013 volume:283 issue:23 firstpage:4370 lastpage:4385 numberofpages:16 journal:THE FEBS JOURNAL http://hdl.handle.net/2108/181089 doi:10.1111/febs.13929 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85000542405 |
op_doi |
https://doi.org/10.1111/febs.13929 |
container_title |
The FEBS Journal |
container_volume |
283 |
container_issue |
23 |
container_start_page |
4370 |
op_container_end_page |
4385 |
_version_ |
1792041662731517952 |