Pyk

• 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	ytzA, pfkA
Get the DNA and protein sequences (Barbe et al., 2009)
Genetic context This image was kindly provided by SubtiList

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

• Sigma factor:

• Regulation:
  • twofold induced by glucose PubMed

• Regulatory mechanism:

• Additional information:

Biological materials

• Mutant:
• GP589 (pyk::cat), available in Stülke lab
• GP600 (pyk::erm), 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