Difference between revisions of "CitZ"

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(Biological materials)
(Biological materials)
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** pGP1776 (for expression, purification in ''E. coli'' with N-terminal Strep-tag, in [[pGP172]], available in [[Jörg Stülke]]'s lab)
 
** pGP1776 (for expression, purification in ''E. coli'' with N-terminal Strep-tag, in [[pGP172]], available in [[Jörg Stülke]]'s lab)
 
** pGP1761 (expression with N-terminal His-tag from ''E. coli'', in [[pWH844]]), available in [[Jörg Stülke]]'s lab
 
** pGP1761 (expression with N-terminal His-tag from ''E. coli'', in [[pWH844]]), available in [[Jörg Stülke]]'s lab
 +
** pGP2515 (N-terminal Strep-tag, purification from ''E. coli'', in [[pGP172]]), available in [[Jörg Stülke]]'s lab
  
 
* '''lacZ fusion:'''
 
* '''lacZ fusion:'''

Revision as of 14:46, 8 December 2014

  • Description: citrate synthase

Gene name citZ
Synonyms citA2
Essential no
Product citrate synthase II
Function TCA cycle
Gene expression levels in SubtiExpress: citZ
Interactions involving this protein in SubtInteract: CitZ
Metabolic function and regulation of this protein in SubtiPathways:
citZ
MW, pI 41 kDa, 5.451
Gene length, protein length 1116 bp, 372 aa
Immediate neighbours icd, ytwI
Sequences Protein DNA DNA_with_flanks
Genetic context
CitZ context.gif
This image was kindly provided by SubtiList
Expression at a glance   PubMed
CitZ expression.png















Categories containing this gene/protein

carbon core metabolism, phosphoproteins

This gene is a member of the following regulons

CcpA regulon, CcpC regulon

The gene

Basic information

  • Locus tag: BSU29140

Phenotypes of a mutant

  • glutamate auxotrophy and a defect in sporulation PubMed

Database entries

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

Additional information

The protein

Basic information/ Evolution

  • Catalyzed reaction/ biological activity: Acetyl-CoA + H2O + oxaloacetate = citrate + CoA (according to Swiss-Prot)
  • Protein family: citrate synthase family (according to Swiss-Prot)

Extended information on the protein

  • Kinetic information:
    • Michaelis-Menten (Random Sequential Reaction Mechanism) PubMed
    • specific activity: 0.141 µmol min-1 (mg protein)-1 PubMed
  • Modification: phosphorylation on Ser-284 PubMed
  • Effectors of protein activity:
    • Inhibited by acetyl-CoA, 2-oxoglutarate and NADH PubMed FEBS Letters
    • Inhibited by citrate and CoA (competitively against acetyl-CoA and non-competitively against oxaloacetate) PubMed
    • Inhibited by ATP competitively in B. subtilis strain 168 and HS 1A17 PubMed PubMed
      • In B. subtilis strain HS 2A2, ATP inhibits a non-competitive fashion PubMed
    • Activated by AMP PubMed

Database entries

  • KEGG entry: [3]

Additional information

  • extensive information on the structure and enzymatic properties of CitZ can be found at Proteopedia

Expression and regulation

  • Regulation:
    • repression by glucose (6.7-fold) (CcpA) PubMed
    • repression by glucose + glutamate (CcpC) PubMed
    • reduced expression at excess citrate concentrations or iron depletion (CitB) PubMed
  • Regulatory mechanism:
    • CcpA: transcription repression, CcpC: transcription repression PubMed
    • CcpC: transcription repression (molecular inducer: citrate) PubMed
    • CitB: mRNA destabilization upon citrate accumulation or iron limitation PubMed
  • Additional information:
    • The mRNA has a long 5' leader region. This may indicate RNA-based regulation PubMed
    • number of protein molecules per cell (minimal medium with glucose and ammonium): 8373 PubMed
    • number of protein molecules per cell (complex medium with amino acids, without glucose): 20578 PubMed
    • number of protein molecules per cell (minimal medium with glucose and ammonium, exponential phase): 22342 PubMed
    • number of protein molecules per cell (minimal medium with glucose and ammonium, early stationary phase after glucose exhaustion): 10224 PubMed
    • number of protein molecules per cell (minimal medium with glucose and ammonium, late stationary phase after glucose exhaustion): 18693 PubMed

Biological materials

  • Expression vector:
    • pGP1120 (N-terminal Strep-tag, for SPINE, purification from B. subtilis, in pGP380) (available in Jörg Stülke's lab)
    • pGP1776 (for expression, purification in E. coli with N-terminal Strep-tag, in pGP172, available in Jörg Stülke's lab)
    • pGP1761 (expression with N-terminal His-tag from E. coli, in pWH844), available in Jörg Stülke's lab
    • pGP2515 (N-terminal Strep-tag, purification from E. coli, in pGP172), 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

Your additional remarks

References

Reviews

Original publications