Artículo

Gil, M.; Lima, A.; Rivera, B.; Rossello, J.; Urdániz, E.; Cascioferro, A.; Carrión, F.; Wehenkel, A.; Bellinzoni, M.; Batthyány, C.; Pritsch, O.; Denicola, A.; Alvarez, M.N.; Carvalho, P.C.; Lisa, M.-N.; Brosch, R.; Piuri, M.; Alzari, P.M.; Durán, R."New substrates and interactors of the mycobacterial Serine/Threonine protein kinase PknG identified by a tailored interactomic approach" (2019) Journal of Proteomics. 192:321-333
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Abstract:

PknG from Mycobacterium tuberculosis is a multidomain Serine/Threonine protein kinase that regulates bacterial metabolism as well as the pathogen's ability to survive inside the host by still uncertain mechanisms. To uncover PknG interactome we developed an affinity purification-mass spectrometry strategy to stepwise recover PknG substrates and interactors; and to identify those involving PknG autophosphorylated docking sites. We report a confident list of 7 new putative substrates and 66 direct or indirect partners indicating that PknG regulates many physiological processes, such as nitrogen and energy metabolism, cell wall synthesis and protein translation. GarA and the 50S ribosomal protein L13, two previously reported substrates of PknG, were recovered in our interactome. Comparative proteome analyses of wild type and pknG null mutant M. tuberculosis strains provided evidence that two kinase interactors, the FHA-domain containing protein GarA and the enzyme glutamine synthetase, are indeed endogenous substrates of PknG, stressing the role of this kinase in the regulation of nitrogen metabolism. Interestingly, a second FHA protein was identified as a PknG substrate. Our results show that PknG phosphorylates specific residues in both glutamine synthetase and FhaA in vitro, and suggest that these proteins are phosphorylated by PknG in living mycobacteria. © 2018 Elsevier B.V.

Registro:

Documento: Artículo
Título:New substrates and interactors of the mycobacterial Serine/Threonine protein kinase PknG identified by a tailored interactomic approach
Autor:Gil, M.; Lima, A.; Rivera, B.; Rossello, J.; Urdániz, E.; Cascioferro, A.; Carrión, F.; Wehenkel, A.; Bellinzoni, M.; Batthyány, C.; Pritsch, O.; Denicola, A.; Alvarez, M.N.; Carvalho, P.C.; Lisa, M.-N.; Brosch, R.; Piuri, M.; Alzari, P.M.; Durán, R.
Filiación:Unidad de Bioquímica y Proteómica Analíticas Institut Pasteur de Montevideo, Instituto de Investigaciones Biológicas Clemente Estable, Mataojo 2020, Montevideo, 11400, Uruguay
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina
Integrated Mycobacterial Pathogenomics Unit, Institut Pasteur, Paris, France
Institut Pasteur de Montevideo, Unidad de Biofísica de Proteínas, Uruguay
Unité de Microbiologie Structurale, Institut Pasteur, CNRS URA 2185, Paris, France
Laboratorio de Fisicoquímica Biológica, Facultad de Ciencias, Universidad de la República, Uruguay
Departamento de Bioquímica, Facultad de Medicina, CEINBIO, Universidad de la República, Uruguay
Laboratory for Proteomics and Protein Engineering, Carlos Chagas Institute, Paraná, Brazil
Unit of Dynamics of Host-Pathogen Interactions, Institut Pasteur, Paris, France
Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, S2002LRK, Rosario, Argentina
Palabras clave:Affinity purification-mass spectrometry; FhaA; glutamine synthetase; Mycobacterium tuberculosis; PknG; Serine/Threonine protein kinase; 50s ribosomal protein L13; 60 kda chaperonin 1; acetyl propionyl coenzyme A carboxylase alpha chain; alcohol dehydrogenase iron containing protein; bacterial enzyme; fhaa protein; gara protein; glutamate ammonia ligase; inorganic pyrophosphatase; pkng protein; protein DnaK; protein serine threonine kinase; unclassified drug; affinity chromatography; Article; autophosphorylation; bacterial cell wall; bacterial strain; controlled study; enzyme analysis; enzyme phosphorylation; enzyme purification; enzyme substrate; ferredoxin iron sulfur binding protein; in vitro study; mass spectrometry; molecular docking; Mycobacterium tuberculosis; nitrogen fixation; nonhuman; priority journal; translation regulation
Año:2019
Volumen:192
Página de inicio:321
Página de fin:333
DOI: http://dx.doi.org/10.1016/j.jprot.2018.09.013
Handle:http://hdl.handle.net/20.500.12110/paper_18743919_v192_n_p321_Gil
Título revista:Journal of Proteomics
Título revista abreviado:J. Proteomics
ISSN:18743919
CAS:glutamate ammonia ligase, 9023-70-5; inorganic pyrophosphatase, 9024-82-2, 9033-44-7; protein serine threonine kinase
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_18743919_v192_n_p321_Gil

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Citas:

---------- APA ----------
Gil, M., Lima, A., Rivera, B., Rossello, J., Urdániz, E., Cascioferro, A., Carrión, F.,..., Durán, R. (2019) . New substrates and interactors of the mycobacterial Serine/Threonine protein kinase PknG identified by a tailored interactomic approach. Journal of Proteomics, 192, 321-333.
http://dx.doi.org/10.1016/j.jprot.2018.09.013
---------- CHICAGO ----------
Gil, M., Lima, A., Rivera, B., Rossello, J., Urdániz, E., Cascioferro, A., et al. "New substrates and interactors of the mycobacterial Serine/Threonine protein kinase PknG identified by a tailored interactomic approach" . Journal of Proteomics 192 (2019) : 321-333.
http://dx.doi.org/10.1016/j.jprot.2018.09.013
---------- MLA ----------
Gil, M., Lima, A., Rivera, B., Rossello, J., Urdániz, E., Cascioferro, A., et al. "New substrates and interactors of the mycobacterial Serine/Threonine protein kinase PknG identified by a tailored interactomic approach" . Journal of Proteomics, vol. 192, 2019, pp. 321-333.
http://dx.doi.org/10.1016/j.jprot.2018.09.013
---------- VANCOUVER ----------
Gil, M., Lima, A., Rivera, B., Rossello, J., Urdániz, E., Cascioferro, A., et al. New substrates and interactors of the mycobacterial Serine/Threonine protein kinase PknG identified by a tailored interactomic approach. J. Proteomics. 2019;192:321-333.
http://dx.doi.org/10.1016/j.jprot.2018.09.013