Pyk

• 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: pyk
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 This image was kindly provided by SubtiList
Expression at a glance   PubMed

Categories containing this gene/protein

ATP synthesis, carbon core metabolism, phosphoproteins

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

• Interactions:

• Localization: cytoplasm PubMed

Database entries

• Structure: 2E28 (Geobacillus stearothermophilus)

• UniProt: P80885

• KEGG entry: [3]

• E.C. number: 2.7.1.40

Additional information

The enzyme is a tetramer with four active sites PubMed

Expression and regulation

• Operon: pfkA-pyk-ytzA PubMed

• Expression browser: pyk PubMed

• Sigma factor:

• Regulation:
  • twofold induced by glucose PubMed

• Regulatory mechanism:

• Additional information:

Biological materials

• Mutant:
• GP589 (pyk::cat), available in Jörg Stülke's lab, PubMed
• GP600 (pyk::erm), available in Jörg Stülke's lab, PubMed

• 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

References

Cuauhtemoc Licona-Cassani, Alvaro R Lara, Natividad Cabrera-Valladares, Adelfo Escalante, Georgina Hernández-Chávez, Alfredo Martinez, Francisco Bolívar, Guillermo Gosset
Inactivation of pyruvate kinase or the phosphoenolpyruvate: sugar phosphotransferase system increases shikimic and dehydroshikimic acid yields from glucose in Bacillus subtilis.
J Mol Microbiol Biotechnol: 2014, 24(1);37-45
[PubMed:24158146] [WorldCat.org] [DOI] (I p)

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)