Difference between revisions of "Mdh"

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(Biological materials)
(Biological materials)
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** pGP1123 (N-terminal Strep-tag, for [[SPINE]], purification from ''B. subtilis'', in [[pGP380]]) (available in [[Stülke]] lab)
 
** pGP1123 (N-terminal Strep-tag, for [[SPINE]], purification from ''B. subtilis'', in [[pGP380]]) (available in [[Stülke]] lab)
 
** for expression, purification in ''E. coli'' with N-terminal His-tag, in [[pWH844]]: pGP385, available in [[Stülke]] lab
 
** for expression, purification in ''E. coli'' with N-terminal His-tag, in [[pWH844]]: pGP385, available in [[Stülke]] lab
** pGP1764 (for expression, purification in ''E. coli'' with N-terminal Strep-tag, in [[pGP172]], available in [[Stülke]] lab)
 
 
** pGP1755 (expression / purification of Mdh-S149A, with N-terminal His-tag from ''E. coli'', in [[pWH844]]), available in [[Stülke]] lab
 
** pGP1755 (expression / purification of Mdh-S149A, with N-terminal His-tag from ''E. coli'', in [[pWH844]]), available in [[Stülke]] lab
 
+
** pGP1764 (for expression, purification in ''E. coli'' with N-terminal Strep-tag, in [[pGP172]], available in [[Stülke]] lab)
 +
 
 
* '''lacZ fusion:'''
 
* '''lacZ fusion:'''
  

Revision as of 15:12, 4 November 2010

  • Description: malate dehydrogenase

Gene name mdh
Synonyms citH
Essential no
Product malate dehydrogenase
Function TCA cycle
Metabolic function and regulation of this protein in SubtiPathways:
Central C-metabolism
MW, pI 33 kDa, 4.727
Gene length, protein length 936 bp, 312 aa
Immediate neighbours phoP, icd
Get the DNA and protein sequences
(Barbe et al., 2009)
Genetic context
Mdh context.gif
This image was kindly provided by SubtiList







The gene

Basic information

  • Locus tag: BSU29120

Phenotypes of a mutant

Database entries

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

Additional information

The protein

Basic information/ Evolution

  • Catalyzed reaction/ biological activity: (S)-malate + NAD+ = oxaloacetate + NADH (according to Swiss-Prot)
  • Protein family: MDH type 3 family (according to Swiss-Prot)
  • Paralogous protein(s):

Extended information on the protein

  • Kinetic information: Reversible Michaelis-Menten PubMed
  • Domains:
  • Modification: phosphorylation on Ser-149 PubMed
  • Cofactor(s):
  • Effectors of protein activity:
    • Inhibited by Mg2+, Ca2+, Zn2+, Ag2+ and Hg2+ PubMed PubMed
    • Inhibited by oxaloacetate (above 1mM) and malate (above 7,7mM) PubMed
  • Localization: cytoplasm (according to Swiss-Prot), membrane associated PubMed

Database entries

  • Structure: 1EMD (E.coli)
  • KEGG entry: [3]

Additional information

The enzyme is a tetramer PubMed

Expression and regulation

  • Regulatory mechanism:
    • CcpA: transcription repression, CcpC: transcription repression PubMed
    • CcpC: transcription repression (molecular inducer: citrate) PubMed
  • Additional information:

Biological materials

  • Mutant: GP719 (spc), available in Stülke lab
  • Expression vector:
    • pGP1123 (N-terminal Strep-tag, for SPINE, purification from B. subtilis, in pGP380) (available in Stülke lab)
    • for expression, purification in E. coli with N-terminal His-tag, in pWH844: pGP385, available in Stülke lab
    • pGP1755 (expression / purification of Mdh-S149A, with N-terminal His-tag from E. coli, in pWH844), available in Stülke lab
    • pGP1764 (for expression, purification in E. coli with N-terminal Strep-tag, in pGP172, available in Stülke lab)
  • lacZ fusion:
  • GFP fusion:
  • two-hybrid system: B. pertussis adenylate cyclase-based bacterial two hybrid system (BACTH), available in Stülke lab
  • Antibody:

Labs working on this gene/protein

Your additional remarks

References

Frederik M Meyer, Jan Gerwig, Elke Hammer, Christina Herzberg, Fabian M Commichau, Uwe Völker, Jörg Stülke
Physical interactions between tricarboxylic acid cycle enzymes in Bacillus subtilis: evidence for a metabolon.
Metab. Eng.: 2011, 13(1);18-27
[PubMed:20933603] [WorldCat.org] [DOI] (I p)

Hannes Hahne, Susanne Wolff, Michael Hecker, Dörte Becher
From complementarity to comprehensiveness--targeting the membrane proteome of growing Bacillus subtilis by divergent approaches.
Proteomics: 2008, 8(19);4123-36
[PubMed:18763711] [WorldCat.org] [DOI] (I p)

Boris Macek, Ivan Mijakovic, Jesper V Olsen, Florian Gnad, Chanchal Kumar, Peter R Jensen, Matthias Mann
The serine/threonine/tyrosine phosphoproteome of the model bacterium Bacillus subtilis.
Mol. Cell Proteomics: 2007, 6(4);697-707
[PubMed:17218307] [WorldCat.org] [DOI] (P p)

Hyun-Jin Kim, Agnes Roux, Abraham L Sonenshein
Direct and indirect roles of CcpA in regulation of Bacillus subtilis Krebs cycle genes.
Mol. Microbiol.: 2002, 45(1);179-90
[PubMed:12100558] [WorldCat.org] (P p)

C Jourlin-Castelli, N Mani, M M Nakano, A L Sonenshein
CcpC, a novel regulator of the LysR family required for glucose repression of the citB gene in Bacillus subtilis.
J. Mol. Biol.: 2000, 295(4);865-78
[PubMed:10656796] [WorldCat.org] [DOI] (P p)

S Jin, M De Jesús-Berríos, A L Sonenshein
A Bacillus subtilis malate dehydrogenase gene.
J. Bacteriol.: 1996, 178(2);560-3
[PubMed:8550482] [WorldCat.org] [DOI] (P p)

S Jin, A L Sonenshein
Transcriptional regulation of Bacillus subtilis citrate synthase genes.
J. Bacteriol.: 1994, 176(15);4680-90
[PubMed:8045899] [WorldCat.org] [DOI] (P p)

A K Tyagi, F A Siddiqui, T A Venkitasubramanian
Studies on the purification and characterization of malate dehydrogenase from Mycobacterium phlei.
Biochim. Biophys. Acta: 1977, 485(2);255-67
[PubMed:922015] [WorldCat.org] [DOI] (P p)

A YOSHIDA
ENZYMIC PROPERTIES OF MALATE DEHYDROGENASE OF BACILLUS SUBTILIS.
J. Biol. Chem.: 1965, 240;1118-24
[PubMed:14284712] [WorldCat.org] (P p)