title: Allosteric regulation of the partitioning of glucose-1-phosphate between glycogen and trehalose biosynthesis in Mycobacterium tuberculosis.
abstract: BACKGROUND
  Mycobacterium tuberculosis is a pathogenic prokaryote adapted to survive in hostile environments. In this organism and other Gram-positive actinobacteria, the metabolic pathways of glycogen and trehalose are interconnected.
  
  
  RESULTS
  In this work we show the production, purification and characterization of recombinant enzymes involved in the partitioning of glucose-1-phosphate between glycogen and trehalose in M. tuberculosis H37Rv, namely: ADP-glucose pyrophosphorylase, glycogen synthase, UDP-glucose pyrophosphorylase and trehalose-6-phosphate synthase. The substrate specificity, kinetic parameters and allosteric regulation of each enzyme were determined. ADP-glucose pyrophosphorylase was highly specific for ADP-glucose while trehalose-6-phosphate synthase used not only ADP-glucose but also UDP-glucose, albeit to a lesser extent. ADP-glucose pyrophosphorylase was allosterically activated primarily by phosphoenolpyruvate and glucose-6-phosphate, while the activity of trehalose-6-phosphate synthase was increased up to 2-fold by fructose-6-phosphate. None of the other two enzymes tested exhibited allosteric regulation.
  
  
  CONCLUSIONS
  Results give information about how the glucose-1-phosphate/ADP-glucose node is controlled after kinetic and regulatory properties of key enzymes for mycobacteria metabolism.
  
  
  CONCLUSIONS
  This work increases our understanding of oligo and polysaccharides metabolism in M. tuberculosis and reinforces the importance of the interconnection between glycogen and trehalose biosynthesis in this human pathogen.
authors.affiliation: Instituto de Agrobiotecnología del Litoral (UNL-CONICET), Facultad de Bioquímica y Ciencias Biológicas, Paraje El Pozo, S3000ZAA Santa Fe, Argentina.
authors.forename: Matías D
authors.initials: MD
authors.lastname: Asención Diez
authors.affiliation: Instituto de Agrobiotecnología del Litoral (UNL-CONICET), Facultad de Bioquímica y Ciencias Biológicas, Paraje El Pozo, S3000ZAA Santa Fe, Argentina.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Demonte
authors.affiliation: Department of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, United Kingdom.
authors.forename: Karl
authors.initials: K
authors.lastname: Syson
authors.affiliation: Instituto de Agrobiotecnología del Litoral (UNL-CONICET), Facultad de Bioquímica y Ciencias Biológicas, Paraje El Pozo, S3000ZAA Santa Fe, Argentina.
authors.forename: Diego G
authors.initials: DG
authors.lastname: Arias
authors.affiliation: Department of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, United Kingdom.
authors.forename: Andrii
authors.initials: A
authors.lastname: Gorelik
authors.affiliation: Instituto de Agrobiotecnología del Litoral (UNL-CONICET), Facultad de Bioquímica y Ciencias Biológicas, Paraje El Pozo, S3000ZAA Santa Fe, Argentina.
authors.forename: Sergio A
authors.initials: SA
authors.lastname: Guerrero
authors.affiliation: Department of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, United Kingdom.
authors.forename: Stephen
authors.initials: S
authors.lastname: Bornemann
authors.affiliation: Instituto de Agrobiotecnología del Litoral (UNL-CONICET), Facultad de Bioquímica y Ciencias Biológicas, Paraje El Pozo, S3000ZAA Santa Fe, Argentina. Electronic address: iglesias@fbcb.unl.edu.ar.
authors.forename: Alberto A
authors.initials: AA
authors.lastname: Iglesias
mesh_terms: D000494:Allosteric Regulation; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D051599:Glucose-1-Phosphate Adenylyltransferase / Q000235:genetics / Q000378:metabolism; D019298:Glucose-6-Phosphate / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism*; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism; D006003:Glycogen / Q000096:biosynthesis*; D006006:Glycogen Synthase / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D053858:Metabolic Networks and Pathways; D008954:Models, Biological; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014199:Trehalose / Q000096:biosynthesis*; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D005958:Glucosephosphates; D011994:Recombinant Proteins; D019298:Glucose-6-Phosphate; D006003:Glycogen; D014199:Trehalose; C031590:glucose-1-phosphate; D005964:Glucosyltransferases; D006006:Glycogen Synthase; C023180:trehalose-6-phosphate synthase; D051599:Glucose-1-Phosphate Adenylyltransferase; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase
keywords: ADP-glucose pyrophosphorylase; Glucose-6-phosphate; Glycogen synthase; Phosphoenolpyruvate; Trehalose-6-phosphate synthase; UDP-glucose pyrophosphorylase