Difference between revisions of "Mdh"

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

Revision as of 09:45, 2 November 2012

  • Description: malate dehydrogenase

Gene name mdh
Synonyms citH
Essential no
Product malate dehydrogenase
Function TCA cycle
Gene expression levels in SubtiExpress: mdh
Interactions involving this protein in SubtInteract: Mdh
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
Expression at a glance   PubMed
Mdh expression.png















Categories containing this gene/protein

carbon core metabolism, membrane proteins, phosphoproteins

This gene is a member of the following regulons

CcpA regulon, CcpC regulon

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:
    • phosphorylated on Arg-156 PubMed
    • 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

Database entries

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

Additional information

  • The enzyme is a tetramer PubMed
  • extensive information on the structure and enzymatic properties of Mdh can be found at Proteopedia

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) & GP1150(spc), available in Stülke lab
  • Expression vector:
    • pGP1123 (N-terminal Strep-tag, for SPINE, purification from B. subtilis, in pGP380) (available in Jörg Stülke's lab)
    • pGP1755 (expression / purification of Mdh-S149A, with N-terminal His-tag from E. coli, in pWH844), available in Jörg Stülke's lab
    • pGP1764 (for expression, purification in E. coli with N-terminal Strep-tag, in pGP172, available in Jörg Stülke's lab)
    • GP1438(mdh-Strep (spc)) & GP1440(mdh-Strep (cat)), purification from B. subtilis, for SPINE, available in Stülke lab
  • lacZ fusion:
  • GFP fusion: GP1431 (spc, based on pGP1870), available in the Stülke lab
  • YFP fusion: GP1429 (spc, based on pGP1871), available in the Stülke lab
  • two-hybrid system: B. pertussis adenylate cyclase-based bacterial two hybrid system (BACTH), available in Jörg Stülke's lab
  • Antibody:

Labs working on this gene/protein

Your additional remarks

References

Alexander K W Elsholz, Kürsad Turgay, Stephan Michalik, Bernd Hessling, Katrin Gronau, Dan Oertel, Ulrike Mäder, Jörg Bernhardt, Dörte Becher, Michael Hecker, Ulf Gerth
Global impact of protein arginine phosphorylation on the physiology of Bacillus subtilis.
Proc. Natl. Acad. Sci. U.S.A.: 2012, 109(19);7451-6
[PubMed:22517742] [WorldCat.org] [DOI] (I p)

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)