Functional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.

The methionine salvage pathway is widely distributed among some eubacteria, yeast, plants and animals and recycles the sulfur-containing metabolite 5-methylthioadenosine (MTA) to methionine. In eukaryotic cells, the methionine salvage pathway takes place in the cytosol and usually involves six enzym...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Camille Mary, Paula Duek, Lisa Salleron, Petra Tienz, Dirk Bumann, Amos Bairoch, Lydie Lane
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b5ef6f0cf1f045ad990f6106fc8b6b7c
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:b5ef6f0cf1f045ad990f6106fc8b6b7c
record_format	dspace
spelling	oai:doaj.org-article:b5ef6f0cf1f045ad990f6106fc8b6b7c2021-11-18T08:03:17ZFunctional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.1932-620310.1371/journal.pone.0052877https://doaj.org/article/b5ef6f0cf1f045ad990f6106fc8b6b7c2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23285211/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The methionine salvage pathway is widely distributed among some eubacteria, yeast, plants and animals and recycles the sulfur-containing metabolite 5-methylthioadenosine (MTA) to methionine. In eukaryotic cells, the methionine salvage pathway takes place in the cytosol and usually involves six enzymatic activities: MTA phosphorylase (MTAP, EC 2.4.2.28), 5'-methylthioribose-1-phosphate isomerase (mtnA, EC 5.3.1.23), 5'-methylthioribulose-1-phosphate dehydratase (mtnB, EC: 4.2.1.109), 2,3-dioxomethiopentane-1-phosphate enolase/phosphatase (mtnC, EC 3.1.3.77), aci-reductone dioxygenase (mtnD, EC 1.13.11.54) and 4-methylthio-2-oxo-butanoate (MTOB) transaminase (EC 2.6.1.-). The aim of this study was to complete the available information on the methionine salvage pathway in human by identifying the enzyme responsible for the dehydratase step. Using a bioinformatics approach, we propose that a protein called APIP could perform this role. The involvement of this protein in the methionine salvage pathway was investigated directly in HeLa cells by transient and stable short hairpin RNA interference. We show that APIP depletion specifically impaired the capacity of cells to grow in media where methionine is replaced by MTA. Using a Shigella mutant auxotroph for methionine, we confirm that the knockdown of APIP specifically affects the recycling of methionine. We also show that mutation of three potential phosphorylation sites does not affect APIP activity whereas mutation of the potential zinc binding site completely abrogates it. Finally, we show that the N-terminal region of APIP that is missing in the short isoform is required for activity. Together, these results confirm the involvement of APIP in the methionine salvage pathway, which plays a key role in many biological functions like cancer, apoptosis, microbial proliferation and inflammation.Camille MaryPaula DuekLisa SalleronPetra TienzDirk BumannAmos BairochLydie LanePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e52877 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Camille Mary Paula Duek Lisa Salleron Petra Tienz Dirk Bumann Amos Bairoch Lydie Lane Functional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.
description	The methionine salvage pathway is widely distributed among some eubacteria, yeast, plants and animals and recycles the sulfur-containing metabolite 5-methylthioadenosine (MTA) to methionine. In eukaryotic cells, the methionine salvage pathway takes place in the cytosol and usually involves six enzymatic activities: MTA phosphorylase (MTAP, EC 2.4.2.28), 5'-methylthioribose-1-phosphate isomerase (mtnA, EC 5.3.1.23), 5'-methylthioribulose-1-phosphate dehydratase (mtnB, EC: 4.2.1.109), 2,3-dioxomethiopentane-1-phosphate enolase/phosphatase (mtnC, EC 3.1.3.77), aci-reductone dioxygenase (mtnD, EC 1.13.11.54) and 4-methylthio-2-oxo-butanoate (MTOB) transaminase (EC 2.6.1.-). The aim of this study was to complete the available information on the methionine salvage pathway in human by identifying the enzyme responsible for the dehydratase step. Using a bioinformatics approach, we propose that a protein called APIP could perform this role. The involvement of this protein in the methionine salvage pathway was investigated directly in HeLa cells by transient and stable short hairpin RNA interference. We show that APIP depletion specifically impaired the capacity of cells to grow in media where methionine is replaced by MTA. Using a Shigella mutant auxotroph for methionine, we confirm that the knockdown of APIP specifically affects the recycling of methionine. We also show that mutation of three potential phosphorylation sites does not affect APIP activity whereas mutation of the potential zinc binding site completely abrogates it. Finally, we show that the N-terminal region of APIP that is missing in the short isoform is required for activity. Together, these results confirm the involvement of APIP in the methionine salvage pathway, which plays a key role in many biological functions like cancer, apoptosis, microbial proliferation and inflammation.
format	article
author	Camille Mary Paula Duek Lisa Salleron Petra Tienz Dirk Bumann Amos Bairoch Lydie Lane
author_facet	Camille Mary Paula Duek Lisa Salleron Petra Tienz Dirk Bumann Amos Bairoch Lydie Lane
author_sort	Camille Mary
title	Functional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.
title_short	Functional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.
title_full	Functional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.
title_fullStr	Functional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.
title_full_unstemmed	Functional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.
title_sort	functional identification of apip as human mtnb, a key enzyme in the methionine salvage pathway.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/b5ef6f0cf1f045ad990f6106fc8b6b7c
work_keys_str_mv	AT camillemary functionalidentificationofapipashumanmtnbakeyenzymeinthemethioninesalvagepathway AT pauladuek functionalidentificationofapipashumanmtnbakeyenzymeinthemethioninesalvagepathway AT lisasalleron functionalidentificationofapipashumanmtnbakeyenzymeinthemethioninesalvagepathway AT petratienz functionalidentificationofapipashumanmtnbakeyenzymeinthemethioninesalvagepathway AT dirkbumann functionalidentificationofapipashumanmtnbakeyenzymeinthemethioninesalvagepathway AT amosbairoch functionalidentificationofapipashumanmtnbakeyenzymeinthemethioninesalvagepathway AT lydielane functionalidentificationofapipashumanmtnbakeyenzymeinthemethioninesalvagepathway
_version_	1718422627290185728

Functional identification of APIP as human mtnB, a key enzyme in the methionine salvage pathway.

Ejemplares similares