Difference between revisions of "RocG"

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(Enzymatic activity of RocG)
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==Enzymatic activity of RocG==
 
==Enzymatic activity of RocG==
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==Function in the control of [[GltC]] activity==
 
==Function in the control of [[GltC]] activity==
 
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<pubmed>15150225,17994626 ,17608797 17183217 20630473  </pubmed>

Revision as of 12:58, 3 March 2015

  • Description: trigger enzyme: catabolic glutamate dehydrogenase induced by arginine, ornithine or proline, subject to carbon catabolite repression

Gene name rocG
Synonyms
Essential no
Product trigger enzyme: glutamate dehydrogenase (major)
Function arginine utilization, controls the activity of GltC
Gene expression levels in SubtiExpress: rocG
Interactions involving this protein in SubtInteract: RocG
Metabolic function and regulation of this protein in SubtiPathways:
rocG
MW, pI 46.2 kDa, 6.28
Gene length, protein length 1272 bp, 424 amino acids
Immediate neighbours rocA, sivA
Sequences Protein DNA DNA_with_flanks
Genetic context
RocG context.gif
This image was kindly provided by SubtiList
Expression at a glance   PubMed
RocG expression.png
















Categories containing this gene/protein

utilization of amino acids, glutamate metabolism, transcription factors and their control, trigger enzyme

This gene is a member of the following regulons

AbrB regulon, AhrC regulon, CcpA regulon, RocR regulon, SigL regulon

The gene

Basic information

  • Locus tag: BSU37790

Phenotypes of a mutant

  • Poor growth on complex media such as SP (sporulation medium). No growth in minimal media with arginine as the only carbon source. Rapid accumulation of suppressor mutants (gudB1)
  • sensitive to ß-lactam antibiotics such as cefuroxime and to fosfomycin (suppressed by activation of gudB) due to the downregulation of the SigW regulon PubMed
  • transcription profile of a rocG gudB mutant strain: GEO PubMed

Database entries

  • DBTBS entry: [1]
  • SubtiList entry: [2]

Additional information

The protein

Basic information/ Evolution

  • Catalyzed reaction/ biological activity: L-glutamate + H2O + NAD+ = 2-oxoglutarate + NH3 + NADH + H+ (according to Swiss-Prot), controls the activity of the GltC transcription activator PubMed
  • Protein family: Glu/Leu/Phe/Val dehydrogenases family (according to Swiss-Prot)
  • Paralogous protein(s): GudB

Extended information on the protein

  • Kinetic information: KM [glutamate] = 2.9 mM, KM [ammonium] = 18 mM PubMed
  • Domains:
  • Modification:
  • Cofactor(s): NAD+/NADH + H+
  • Effectors of protein activity:

Database entries

  • Structure: 3K92 (super-repressor mutant that is capable of constitutive inactivation of GltC, E93K mutation) PubMed
  • KEGG entry: [4]

Additional information

Expression and regulation

  • Regulation:
  • Additional information:

Activation by RocR requires binding of RocR to a downstream element PubMed

    • number of protein molecules per cell (complex medium with amino acids, without glucose): 16024 PubMed

Biological materials

  • Mutant: GP747 (spc), GP726 (aphA3), GP810 (del tet), GP1157 (cat) all available in Jörg Stülke's lab
  • Expression vector:
    • expression of native rocG in B. subtilis: pGP529 (in pBQ200), available in Jörg Stülke's lab PubMed
    • for purification of RocG from E. coli carrying an N-terminal Strep-tag: pGP902 (in pGP172), a series of rocG variants is also available in pGP172, available in Jörg Stülke's lab
    • for expression/ purification from E. coli with N-terminal His-tag and thrombin cleavage site, in pWH844: pGP860, available in Jörg Stülke's lab
    • purification from B. subtilis with an N-terminal Strep-tag, for SPINE, (in pGP380): pGP1709, available in Jörg Stülke's lab
  • lacZ fusion:
  • GFP fusion:
  • two-hybrid system: B. pertussis adenylate cyclase-based bacterial two hybrid system (BACTH), available in Jörg Stülke's lab

Labs working on this gene/protein

Linc Sonenshein, Tufts University, Boston, MA, USA Homepage

Jörg Stülke, University of Göttingen, Germany Homepage

Fabian Commichau University of Göttingen, Germany Homepage

Your additional remarks

References

Reviews

Katrin Gunka, Fabian M Commichau
Control of glutamate homeostasis in Bacillus subtilis: a complex interplay between ammonium assimilation, glutamate biosynthesis and degradation.
Mol Microbiol: 2012, 85(2);213-24
[PubMed:22625175] [WorldCat.org] [DOI] (I p)

Jason R Treberg, Margaret E Brosnan, Malcolm Watford, John T Brosnan
On the reversibility of glutamate dehydrogenase and the source of hyperammonemia in the hyperinsulinism/hyperammonemia syndrome.
Adv Enzyme Regul: 2010, 50(1);34-43
[PubMed:19895831] [WorldCat.org] [DOI] (I p)

Victoria I Bunik, Alisdair R Fernie
Metabolic control exerted by the 2-oxoglutarate dehydrogenase reaction: a cross-kingdom comparison of the crossroad between energy production and nitrogen assimilation.
Biochem J: 2009, 422(3);405-21
[PubMed:19698086] [WorldCat.org] [DOI] (I e)

N M Brunhuber, J S Blanchard
The biochemistry and enzymology of amino acid dehydrogenases.
Crit Rev Biochem Mol Biol: 1994, 29(6);415-67
[PubMed:7705101] [WorldCat.org] [DOI] (P p)

R C Hudson, R M Daniel
L-glutamate dehydrogenases: distribution, properties and mechanism.
Comp Biochem Physiol B: 1993, 106(4);767-92
[PubMed:8299344] [WorldCat.org] [DOI] (P p)


Enzymatic activity of RocG


Function in the control of GltC activity

Expression of rocG

Structural analysis of glutamate dehydrogenase

Katrin Gunka, Joseph A Newman, Fabian M Commichau, Christina Herzberg, Cecilia Rodrigues, Lorraine Hewitt, Richard J Lewis, Jörg Stülke
Functional dissection of a trigger enzyme: mutations of the bacillus subtilis glutamate dehydrogenase RocG that affect differentially its catalytic activity and regulatory properties.
J Mol Biol: 2010, 400(4);815-27
[PubMed:20630473] [WorldCat.org] [DOI] (I p)

T J Stillman, P J Baker, K L Britton, D W Rice
Conformational flexibility in glutamate dehydrogenase. Role of water in substrate recognition and catalysis.
J Mol Biol: 1993, 234(4);1131-9
[PubMed:8263917] [WorldCat.org] [DOI] (P p)

Bypass of rocG mutations


Additional publications