NADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.

Mammalian methionine adenosyltransferase II (MAT II) is the only hetero-oligomer in this family of enzymes that synthesize S-adenosylmethionine using methionine and ATP as substrates. Binding of regulatory β subunits and catalytic α2 dimers is known to increase the affinity for methionine, although...

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Autores principales:	Beatriz González, Francisco Garrido, Rebeca Ortega, Marta Martínez-Júlvez, Ainhoa Revilla-Guarinos, Yolanda Pérez-Pertejo, Adrián Velázquez-Campoy, Julia Sanz-Aparicio, María A Pajares
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7b4b669595844ce0a855757355bdd4c1
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spelling	oai:doaj.org-article:7b4b669595844ce0a855757355bdd4c12021-11-18T08:07:34ZNADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.1932-620310.1371/journal.pone.0050329https://doaj.org/article/7b4b669595844ce0a855757355bdd4c12012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23189196/?tool=EBIhttps://doaj.org/toc/1932-6203Mammalian methionine adenosyltransferase II (MAT II) is the only hetero-oligomer in this family of enzymes that synthesize S-adenosylmethionine using methionine and ATP as substrates. Binding of regulatory β subunits and catalytic α2 dimers is known to increase the affinity for methionine, although scarce additional information about this interaction is available. This work reports the use of recombinant α2 and β subunits to produce oligomers showing kinetic parameters comparable to MAT II purified from several tissues. According to isothermal titration calorimetry data and densitometric scanning of the stained hetero-oligomer bands on denatured gels, the composition of these oligomers is that of a hetero-trimer with α2 dimers associated to single β subunits. Additionally, the regulatory subunit is able to bind NADP(+) with a 1:1 stoichiometry, the cofactor enhancing β to α2-dimer binding affinity. Mutants lacking residues involved in NADP(+) binding and N-terminal truncations of the β subunit were able to oligomerize with α2-dimers, although the kinetic properties appeared altered. These data together suggest a role for both parts of the sequence in the regulatory role exerted by the β subunit on catalysis. Moreover, preparation of a structural model for the hetero-oligomer, using the available crystal data, allowed prediction of the regions involved in β to α2-dimer interaction. Finally, the implications that the presence of different N-terminals in the β subunit could have on MAT II behavior are discussed in light of the recent identification of several splicing forms of this subunit in hepatoma cells.Beatriz GonzálezFrancisco GarridoRebeca OrtegaMarta Martínez-JúlvezAinhoa Revilla-GuarinosYolanda Pérez-PertejoAdrián Velázquez-CampoyJulia Sanz-AparicioMaría A PajaresPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 11, p e50329 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Beatriz González Francisco Garrido Rebeca Ortega Marta Martínez-Júlvez Ainhoa Revilla-Guarinos Yolanda Pérez-Pertejo Adrián Velázquez-Campoy Julia Sanz-Aparicio María A Pajares NADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.
description	Mammalian methionine adenosyltransferase II (MAT II) is the only hetero-oligomer in this family of enzymes that synthesize S-adenosylmethionine using methionine and ATP as substrates. Binding of regulatory β subunits and catalytic α2 dimers is known to increase the affinity for methionine, although scarce additional information about this interaction is available. This work reports the use of recombinant α2 and β subunits to produce oligomers showing kinetic parameters comparable to MAT II purified from several tissues. According to isothermal titration calorimetry data and densitometric scanning of the stained hetero-oligomer bands on denatured gels, the composition of these oligomers is that of a hetero-trimer with α2 dimers associated to single β subunits. Additionally, the regulatory subunit is able to bind NADP(+) with a 1:1 stoichiometry, the cofactor enhancing β to α2-dimer binding affinity. Mutants lacking residues involved in NADP(+) binding and N-terminal truncations of the β subunit were able to oligomerize with α2-dimers, although the kinetic properties appeared altered. These data together suggest a role for both parts of the sequence in the regulatory role exerted by the β subunit on catalysis. Moreover, preparation of a structural model for the hetero-oligomer, using the available crystal data, allowed prediction of the regions involved in β to α2-dimer interaction. Finally, the implications that the presence of different N-terminals in the β subunit could have on MAT II behavior are discussed in light of the recent identification of several splicing forms of this subunit in hepatoma cells.
format	article
author	Beatriz González Francisco Garrido Rebeca Ortega Marta Martínez-Júlvez Ainhoa Revilla-Guarinos Yolanda Pérez-Pertejo Adrián Velázquez-Campoy Julia Sanz-Aparicio María A Pajares
author_facet	Beatriz González Francisco Garrido Rebeca Ortega Marta Martínez-Júlvez Ainhoa Revilla-Guarinos Yolanda Pérez-Pertejo Adrián Velázquez-Campoy Julia Sanz-Aparicio María A Pajares
author_sort	Beatriz González
title	NADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.
title_short	NADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.
title_full	NADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.
title_fullStr	NADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.
title_full_unstemmed	NADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.
title_sort	nadp+ binding to the regulatory subunit of methionine adenosyltransferase ii increases intersubunit binding affinity in the hetero-trimer.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/7b4b669595844ce0a855757355bdd4c1
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NADP+ binding to the regulatory subunit of methionine adenosyltransferase II increases intersubunit binding affinity in the hetero-trimer.

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