Difference between revisions of "Mdh"

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(Biological materials)
(Biological materials)
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* '''lacZ fusion:'''
 
* '''lacZ fusion:'''
  
* '''GFP 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
 
* '''two-hybrid system:''' ''B. pertussis'' adenylate cyclase-based bacterial two hybrid system ([[BACTH]]), available in [[Jörg Stülke]]'s lab

Revision as of 08:31, 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)
    • 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
    • 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