Pyk

• Description: pyruvate kinase, glycolytic enzyme
  
  

Gene name	pyk
Synonyms	pykA
Essential	no
Product	pyruvate kinase
Function	catabolic enzyme in glycolysis
MW, pI	61,9 kDa, 4.88
Gene length, protein length	1755 bp, 585 amino acids
Immediate neighbours	pfkA, ytzA
Gene sequence (+200bp)	Protein sequence
Genetic context

The gene

Basic information

• Coordinates: 2983830 - 2985584

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]

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity:ADP + phosphoenolpyruvate --> ATP + pyruvate, the reaction is irreversible under physiological conditions

• Protein family: pyruvate kinase family, (C-terminal section: PEP-utilizing enzyme family)

• Paralogous protein(s):

Extended information on the protein

• Kinetic information:

• Domains:

• Modification: phosphorylation (Ser36 and Ser535 or 546) PubMed

• Cofactor(s): magnesium ion, potassium

• Effectors of protein activity: activated by PEP PubMed

• Interactions:

• Localization: cytoplasm PubMed

Database entries

• Structure: 2E28 (from Geobacillus stearothermophilus) NCBI

• Swiss prot entry: [3]

• KEGG entry: [4]

• E.C. number: [5]

Expression and regulation

• Operon: pfkA pyk PubMed

• Sigma factor:

• Regulation: twofold induced by glucose PubMed

• Regulatory mechanism:

• Additional information:

Biological materials

• Mutant: GP590 (cat), 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

References

1. Ludwig, H., Homuth, G., Schmalisch, M., Dyka, F. M., Hecker, M., and Stülke, J. (2001) Transcription of glycolytic genes and operons in Bacillus subtilis: evidence for the presence of multiple levels of control of the gapA operon. Mol Microbiol 41, 409-422.PubMed
2. Jannière, L., Canceill, D., Suski, C., Kanga, S., Dalmais, B., Lestini, R., Monnier, A. F., Chapuis, J., Bolotin, A., Titok, M., Le Chatelier, E., and Ehrlich, S. D. (2007) Genetic evidence for a link between glycolysis and DNA replication. PLoS ONE 2, e447. PubMed
3. Fry, B., Zhu, T., Domach, M. M., Koepsel, R. R., Phalakornkule, C., and Ataai, M. M. (2000) Characterization of growth and acid formation in a Bacillus subtilis pyruvate kinase mutant. Appl Env Microbiol 66: 4045-4049. PubMed
4. Macek et al. (2007) The serine/ threonine/ tyrosine phosphoproteome of the model bacterium Bacillus subtilis. Mol. Cell. Proteomics 6: 697-707 PubMed