Difference between revisions of "Pyk"

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(Biological materials)
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=Biological materials =
 
=Biological materials =
  
* '''Mutant:''' GP590 (cat), available in [[Stülke]] lab
+
* '''Mutant:''' GP590 (cat), GP510 (spc), available in [[Stülke]] lab
  
 
* '''Expression vector:'''  
 
* '''Expression vector:'''  
Expression in ''E. coli'', N-terminal His-tag: pGP1100 (in [[pWH844]]), available in [[Stülke]] lab
+
** expression in ''E. coli'', N-terminal His-tag: pGP1100 (in [[pWH844]]), available in [[Stülke]] lab
 
+
** expression in ''B. subtilis'', native protein: pGP1411 (in [[pBQ200]]), available in [[Stülke]] lab
Expression in ''B. subtilis'', native protein: pGP1411 (in [[pBQ200]]), available in [[Stülke]] lab
+
** expression in ''B. subtilis'', N-terminal Strep-tag: pGP1409 (in [[pGP380]]), available in [[Stülke]] lab
 
+
** expression in ''B. subtilis'', C-terminal Strep-tag: pGP1410 (in [[pGP382]]), available in [[Stülke]] lab
Expression in ''B. subtilis'', N-terminal Strep-tag: pGP1409 (in [[pGP380]]), available in [[Stülke]] lab
 
 
 
Expression in ''B. subtilis'', C-terminal Strep-tag: pGP1410 (in [[pGP382]]), available in [[Stülke]] lab
 
  
 
* '''lacZ fusion:''' see ''[[pfkA]]''
 
* '''lacZ fusion:''' see ''[[pfkA]]''

Revision as of 16:34, 2 September 2009

  • Description: pyruvate kinase, glycolytic enzyme

Gene name pyk
Synonyms pykA
Essential no
Product pyruvate kinase
Function catabolic enzyme in glycolysis
Metabolic function and regulation of this protein in SubtiPathways:
Central C-metabolism
MW, pI 61,9 kDa, 4.88
Gene length, protein length 1755 bp, 585 amino acids
Immediate neighbours pfkA, ytzA
Get the DNA and protein sequences
(Barbe et al., 2009)
Genetic context
Pyk context.gif
This image was kindly provided by SubtiList







The gene

Basic information

  • Locus tag: BSU29180

Phenotypes of a mutant

Unable to grow with non-PTS carbohydrates (such as glucitol or glycerol) as single carbon source.

Database entries

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

Additional information

The protein

Basic information/ Evolution

  • Catalyzed reaction/ biological activity: ADP + phosphoenolpyruvate --> ATP + pyruvate
    • The reaction is irreversible under physiological conditions
  • Protein family: PEP-utilizing enzyme family (according to Swiss-Prot) pyruvate kinase family, (C-terminal section: PEP-utilizing enzyme family)
  • Paralogous protein(s):

Extended information on the protein

  • Kinetic information: Allosteric Regulation PubMed
  • Domains:
  • Modification: phosphorylation on Ser36 PubMed, PubMed, phosphorylation on Ser536 or Ser546 PubMed, please note that the Seris not on position 536 but rather at 538
  • Cofactor(s): Mg2+, K+
  • Effectors of protein activity:
    • Activated by PEP (Hill Coefficient 1,8) PubMed PubMed
    • Allosterically activated by AMP PubMed
    • Activation by r5p and ADP PubMed
    • Inhibition by ADP and f16bp in high concentrations; and ATP PubMed
  • Interactions:
  • Localization: cytoplasm PubMed

Database entries

  • Structure: 2E28 (Geobacillus stearothermophilus)
  • KEGG entry: [3]

Additional information

The enzyme is a tetramer with four active sites PubMed

Expression and regulation

  • Sigma factor:
  • Regulation: twofold induced by glucose PubMed
  • Regulatory mechanism:
  • Additional information:

Biological materials

  • Mutant: GP590 (cat), GP510 (spc), available in Stülke lab
  • Expression vector:
    • expression in E. coli, N-terminal His-tag: pGP1100 (in pWH844), available in Stülke lab
    • expression in B. subtilis, native protein: pGP1411 (in pBQ200), available in Stülke lab
    • expression in B. subtilis, N-terminal Strep-tag: pGP1409 (in pGP380), available in Stülke lab
    • expression in B. subtilis, C-terminal Strep-tag: pGP1410 (in pGP382), available in Stülke lab
  • lacZ fusion: see pfkA
  • 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

Jörg Stülke, University of Göttingen, Germany Homepage

Your additional remarks

References