Difference between revisions of "Pyk"

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= [[Categories]] containing this gene/protein =
 
= [[Categories]] containing this gene/protein =
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* '''Expression browser:''' [http://genome.jouy.inra.fr/cgi-bin/seb/viewdetail.py?id=pyk_2984788_2986545_-1 pyk] {{PubMed|22383849}}
 
* '''Expression browser:''' [http://genome.jouy.inra.fr/cgi-bin/seb/viewdetail.py?id=pyk_2984788_2986545_-1 pyk] {{PubMed|22383849}}
  
* '''Sigma factor:'''  
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* '''[[Sigma factor]]:'''  
  
 
* '''Regulation:'''  
 
* '''Regulation:'''  
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*  '''Mutant:'''  
 
*  '''Mutant:'''  
* GP589 (''pyk''::''cat''), available in [[Stülke]] lab
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* GP589 (''pyk''::''cat''), available in [[Jörg Stülke]]'s lab, {{PubMed|23420519}}
* GP600 (''pyk''::''erm''), available in [[Stülke]] lab
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* GP600 (''pyk''::''erm''), available in [[Jörg Stülke]]'s lab, {{PubMed|23420519}}
  
 
* '''Expression vector:'''  
 
* '''Expression vector:'''  
** expression in ''E. coli'', N-terminal His-tag: pGP1100 (in [[pWH844]]), available in [[Stülke]] lab
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** expression in ''E. coli'', N-terminal His-tag: pGP1100 (in [[pWH844]]), available in [[Jörg Stülke]]'s lab
** expression in ''B. subtilis'', native protein: pGP1411 (in [[pBQ200]]), available in [[Stülke]] lab
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** expression in ''B. subtilis'', native protein: pGP1411 (in [[pBQ200]]), available in [[Jörg Stülke]]'s lab
** expression in ''B. subtilis'', N-terminal Strep-tag: pGP1409 (in [[pGP380]]), available in [[Stülke]] lab
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** expression in ''B. subtilis'', N-terminal Strep-tag: pGP1409 (in [[pGP380]]), available in [[Jörg Stülke]]'s lab
** expression in ''B. subtilis'', C-terminal Strep-tag: pGP1410 (in [[pGP382]]), available in [[Stülke]] lab
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** expression in ''B. subtilis'', C-terminal Strep-tag: pGP1410 (in [[pGP382]]), available in [[Jörg Stülke]]'s lab
  
 
* '''lacZ fusion:''' see ''[[pfkA]]''
 
* '''lacZ fusion:''' see ''[[pfkA]]''
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* '''GFP fusion:'''
 
* '''GFP fusion:'''
  
* '''two-hybrid system:''' ''B. pertussis'' adenylate cyclase-based bacterial two hybrid system ([[BACTH]]), available in [[Stülke]] lab
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* '''two-hybrid system:''' ''B. pertussis'' adenylate cyclase-based bacterial two hybrid system ([[BACTH]]), available in [[Jörg Stülke]]'s lab
  
 
* '''Antibody:'''
 
* '''Antibody:'''
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=References=
 
=References=
  
<pubmed>17726680 16493705 11489127 17505547 10966427 17218307 3711058 4623707 21821766 22846916 </pubmed>
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<pubmed>17726680 16493705 11489127 17505547 10966427 17218307 3711058 4623707 21821766 22846916 23420519</pubmed>
  
 
[[Category:Protein-coding genes]]
 
[[Category:Protein-coding genes]]

Revision as of 16:02, 13 July 2013

  • Description: pyruvate kinase, glycolytic enzyme

Gene name pyk
Synonyms pykA
Essential no
Product pyruvate kinase
Function catabolic enzyme in glycolysis
Gene expression levels in SubtiExpress: pyk
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 ytzA, pfkA
Sequences Protein DNA DNA_with_flanks
Genetic context
Pyk context.gif
This image was kindly provided by SubtiList
Expression at a glance   PubMed
Pyk expression.png















Categories containing this gene/protein

ATP synthesis, carbon core metabolism, phosphoproteins

This gene is a member of the following regulons

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

  • A mutation was found in this gene after evolution under relaxed selection for sporulation PubMed

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 Ser is 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

Database entries

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

Additional information

The enzyme is a tetramer with four active sites PubMed

Expression and regulation

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

Biological materials

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

Jörg Stülke, University of Göttingen, Germany Homepage

Your additional remarks

References

Fabian M Commichau, Nico Pietack, Jörg Stülke
Essential genes in Bacillus subtilis: a re-evaluation after ten years.
Mol Biosyst: 2013, 9(6);1068-75
[PubMed:23420519] [WorldCat.org] [DOI] (I p)

Natividad Cabrera-Valladares, Luz M Martínez, Noemí Flores, Georgina Hernández-Chávez, Alfredo Martínez, Francisco Bolívar, Guillermo Gosset
Physiologic consequences of glucose transport and phosphoenolpyruvate node modifications in Bacillus subtilis 168.
J Mol Microbiol Biotechnol: 2012, 22(3);177-97
[PubMed:22846916] [WorldCat.org] [DOI] (I p)

Christopher T Brown, Laura K Fishwick, Binna M Chokshi, Marissa A Cuff, Jay M Jackson, Travis Oglesby, Alison T Rioux, Enrique Rodriguez, Gregory S Stupp, Austin H Trupp, James S Woollcombe-Clarke, Tracy N Wright, William J Zaragoza, Jennifer C Drew, Eric W Triplett, Wayne L Nicholson
Whole-genome sequencing and phenotypic analysis of Bacillus subtilis mutants following evolution under conditions of relaxed selection for sporulation.
Appl Environ Microbiol: 2011, 77(19);6867-77
[PubMed:21821766] [WorldCat.org] [DOI] (I p)

Christine Eymann, Dörte Becher, Jörg Bernhardt, Katrin Gronau, Anja Klutzny, Michael Hecker
Dynamics of protein phosphorylation on Ser/Thr/Tyr in Bacillus subtilis.
Proteomics: 2007, 7(19);3509-26
[PubMed:17726680] [WorldCat.org] [DOI] (P p)

Laurent Jannière, Danielle Canceill, Catherine Suski, Sophie Kanga, Bérengère Dalmais, Roxane Lestini, Anne-Françoise Monnier, Jérôme Chapuis, Alexander Bolotin, Marina Titok, Emmanuelle Le Chatelier, S Dusko Ehrlich
Genetic evidence for a link between glycolysis and DNA replication.
PLoS One: 2007, 2(5);e447
[PubMed:17505547] [WorldCat.org] [DOI] (I e)

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)

Alain Lévine, Françoise Vannier, Cédric Absalon, Lauriane Kuhn, Peter Jackson, Elaine Scrivener, Valérie Labas, Joëlle Vinh, Patrick Courtney, Jérôme Garin, Simone J Séror
Analysis of the dynamic Bacillus subtilis Ser/Thr/Tyr phosphoproteome implicated in a wide variety of cellular processes.
Proteomics: 2006, 6(7);2157-73
[PubMed:16493705] [WorldCat.org] [DOI] (P p)

H Ludwig, G Homuth, M Schmalisch, F M Dyka, M Hecker, J Stülke
Transcription of glycolytic genes and operons in Bacillus subtilis: evidence for the presence of multiple levels of control of the gapA operon.
Mol Microbiol: 2001, 41(2);409-22
[PubMed:11489127] [WorldCat.org] [DOI] (P p)

B Fry, T Zhu, M M Domach, R R Koepsel, C Phalakornkule, M M Ataai
Characterization of growth and acid formation in a Bacillus subtilis pyruvate kinase mutant.
Appl Environ Microbiol: 2000, 66(9);4045-9
[PubMed:10966427] [WorldCat.org] [DOI] (P p)

H Sakai, K Suzuki, K Imahori
Purification and properties of pyruvate kinase from Bacillus stearothermophilus.
J Biochem: 1986, 99(4);1157-67
[PubMed:3711058] [WorldCat.org] [DOI] (P p)

M Diesterhaft, E Freese
Pyruvate kinase of bacillus subtilis.
Biochim Biophys Acta: 1972, 268(2);373-80
[PubMed:4623707] [WorldCat.org] [DOI] (P p)