Characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity
The gene encoding MG Orn has been identified from a metagenomic library created from the intertidal zone in Svalbard and encodes a protein of 184 amino acid residues. The mg orn gene has been cloned, recombinantly expressed in Escherichia coli, and purified to homogeneity. Biochemical characterizati...
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ftdoajarticles:oai:doaj.org/article:82adfe6207534dca986f80f83d94409f 2023-05-15T18:29:49+02:00 Characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity Yvonne Piotrowski Kristel Berg David Paul Klebl Ingar Leiros Atle Noralf Larsen 2019-10-01T00:00:00Z https://doi.org/10.1002/2211-5463.12720 https://doaj.org/article/82adfe6207534dca986f80f83d94409f EN eng Wiley https://doi.org/10.1002/2211-5463.12720 https://doaj.org/toc/2211-5463 2211-5463 doi:10.1002/2211-5463.12720 https://doaj.org/article/82adfe6207534dca986f80f83d94409f FEBS Open Bio, Vol 9, Iss 10, Pp 1674-1688 (2019) crystal structure homodimer metagenome nuclease activity oligoribonuclease RNA Biology (General) QH301-705.5 article 2019 ftdoajarticles https://doi.org/10.1002/2211-5463.12720 2022-12-31T03:13:28Z The gene encoding MG Orn has been identified from a metagenomic library created from the intertidal zone in Svalbard and encodes a protein of 184 amino acid residues. The mg orn gene has been cloned, recombinantly expressed in Escherichia coli, and purified to homogeneity. Biochemical characterization of the enzyme showed that it efficiently degrades short RNA oligonucleotide substrates of 2mer to 10mer of length and has an absolute requirement for divalent cations for optimal activity. The enzyme is more heat‐labile than its counterpart from E. coli and exists as a homodimer in solution. The crystal structure of the enzyme has been determined to a resolution of 3.15 Å, indicating an important role of a disulfide bridge for the homodimer formation and as such for the function of MG Orn. Substitution of the Cys110 residue with either Gly or Ala hampered the dimer formation and severely affected the enzyme's ability to act on RNA. A conserved loop containing His128‐Tyr129‐Arg130 in the neighboring monomer is probably involved in efficient binding and processing of longer RNA substrates than diribonucleotides. Article in Journal/Newspaper Svalbard Directory of Open Access Journals: DOAJ Articles Svalbard FEBS Open Bio 9 10 1674 1688 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
crystal structure homodimer metagenome nuclease activity oligoribonuclease RNA Biology (General) QH301-705.5 |
spellingShingle |
crystal structure homodimer metagenome nuclease activity oligoribonuclease RNA Biology (General) QH301-705.5 Yvonne Piotrowski Kristel Berg David Paul Klebl Ingar Leiros Atle Noralf Larsen Characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity |
topic_facet |
crystal structure homodimer metagenome nuclease activity oligoribonuclease RNA Biology (General) QH301-705.5 |
description |
The gene encoding MG Orn has been identified from a metagenomic library created from the intertidal zone in Svalbard and encodes a protein of 184 amino acid residues. The mg orn gene has been cloned, recombinantly expressed in Escherichia coli, and purified to homogeneity. Biochemical characterization of the enzyme showed that it efficiently degrades short RNA oligonucleotide substrates of 2mer to 10mer of length and has an absolute requirement for divalent cations for optimal activity. The enzyme is more heat‐labile than its counterpart from E. coli and exists as a homodimer in solution. The crystal structure of the enzyme has been determined to a resolution of 3.15 Å, indicating an important role of a disulfide bridge for the homodimer formation and as such for the function of MG Orn. Substitution of the Cys110 residue with either Gly or Ala hampered the dimer formation and severely affected the enzyme's ability to act on RNA. A conserved loop containing His128‐Tyr129‐Arg130 in the neighboring monomer is probably involved in efficient binding and processing of longer RNA substrates than diribonucleotides. |
format |
Article in Journal/Newspaper |
author |
Yvonne Piotrowski Kristel Berg David Paul Klebl Ingar Leiros Atle Noralf Larsen |
author_facet |
Yvonne Piotrowski Kristel Berg David Paul Klebl Ingar Leiros Atle Noralf Larsen |
author_sort |
Yvonne Piotrowski |
title |
Characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity |
title_short |
Characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity |
title_full |
Characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity |
title_fullStr |
Characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity |
title_full_unstemmed |
Characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity |
title_sort |
characterization of an intertidal zone metagenome oligoribonuclease and the role of the intermolecular disulfide bond for homodimer formation and nuclease activity |
publisher |
Wiley |
publishDate |
2019 |
url |
https://doi.org/10.1002/2211-5463.12720 https://doaj.org/article/82adfe6207534dca986f80f83d94409f |
geographic |
Svalbard |
geographic_facet |
Svalbard |
genre |
Svalbard |
genre_facet |
Svalbard |
op_source |
FEBS Open Bio, Vol 9, Iss 10, Pp 1674-1688 (2019) |
op_relation |
https://doi.org/10.1002/2211-5463.12720 https://doaj.org/toc/2211-5463 2211-5463 doi:10.1002/2211-5463.12720 https://doaj.org/article/82adfe6207534dca986f80f83d94409f |
op_doi |
https://doi.org/10.1002/2211-5463.12720 |
container_title |
FEBS Open Bio |
container_volume |
9 |
container_issue |
10 |
container_start_page |
1674 |
op_container_end_page |
1688 |
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1766213208841912320 |