pyk
168
[metabolite|pyruvate] kinase, glycolytic enzyme, involved in control of [category|SW.3.1.1|DNA replication]
locus
BSU_29180
Molecular weight
62.00 kDa
pI
4.88
function
catabolic enzyme in glycolysis, control of [category|SW.3.1.1|DNA replication]
product
[metabolite|pyruvate] kinase
essential
no
ec
2.7.1.40
synonyms
pyk, pykA
Outlinks
Genomic Context
Categories containing this gene/protein
List of homologs in different organisms, belongs to COG3848 (Galperin et al., 2021)
This gene is a member of the following regulons
Gene
Coordinates
2,984,788 → 2,986,545
Phenotypes of a mutant
unable to grow with non-[protein|14ED1AF5038F43F3B151FCBABE6CFC5A2DA3AA6E|ptsI] carbohydrates (such as [metabolite|glucitol] or [metabolite|glycerol]) as single carbon source
suppression of ''[gene|41872E2EF00C79918DD077F2EF78F37E24FEB110|ftsZ]''(ts) mutation (reverted by addition of [metabolite|pyruvate]) [Pubmed|24825009]
the mutant cells are thinner and shorter than wild type cells [pubmed|34846166]
The protein
Catalyzed reaction/ biological activity
[metabolite|ADP] + [metabolite|PEP] → [metabolite|ATP] + [metabolite|pyruvate]
The reaction is irreversible under physiological conditions
Protein family
C-terminal part: [wiki|PEP-utilizing enzyme family] (according to UniProt)
pyruvate kinase family (single member, according to UniProt)
[wiki|Domains]
extra C-terminal domain (ECTD): aa 475 - 585 [pubmed|39211278]
[wiki|Cofactors]
Mg2+, K+
Structure
[PDB|2E28] (Geobacillus stearothermophilus) [pubmed|18511452]
[AF|P80885]
Modification
phosphorylation on Ser36 [Pubmed|17218307], [Pubmed|16493705], phosphorylation on Ser536 or Ser546 [Pubmed|17726680], please note that the Ser is not on position 536 but rather at 538
Effectors of protein activity
activated by [metabolite|PEP] (Hill Coefficient 1,8) [Pubmed|4623707] [Pubmed|3711058]
allosterically activated by [metabolite|AMP], this requires the extra C-terminal domain [Pubmed|39211278,3711058]
activation by r5p and [metabolite|ADP], this requires the extra C-terminal domain [Pubmed|39211278,3711058]
inhibition by [metabolite|ADP] and [metabolite|fructose-1,6-bisphosphate] in high concentrations; and [metabolite|ATP], , this requires the extra C-terminal domain [Pubmed|39211278,3711058]
[wiki|Localization]
cytoplasm [Pubmed|16479537]
Additional information
the enzyme is a tetramer with four active sites [Pubmed|39211278,3711058]
belongs to the 100 [http://subtiwiki.uni-goettingen.de/wiki//index.php/Most_abundant_proteins most abundant proteins] [PubMed|15378759]
the enzyme is inhibited under gluconeogenetic conditions [pubmed|39211278]
Expression and Regulation
Operons
genes
[gene|A7E31A5EBCF343B3A841E5F6E9CDE3CA5D1622EB|pfkA]-[gene|F76A03A71DADC32C7166E37B994EFED019FDF8A4|pyk]-[gene|1C6D935B36416327FE63CABC53440441E8563D7E|ytzA]
description
[Pubmed|11489127]
regulation
twofold induced by glucose [Pubmed|11489127]
Biological materials
Mutant
GP589 (''pyk''::''cat''), available in [wiki|Jörg Stülke]'s lab, [Pubmed|23420519]
GP600 (''pyk''::''erm''), available in [wiki|Jörg Stülke]'s lab, [Pubmed|23420519]
GP1745: BSB1 ''pyk::aphA3'', available in [wiki|Jörg Stülke]' lab
BKE29180 (Δ[gene|F76A03A71DADC32C7166E37B994EFED019FDF8A4|pyk]::erm trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKE29180 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CATTTGGTTCACTTCCTTCT, downstream forward: _UP4_TAATTACAGGTGAAAATGGA
BKK29180 (Δ[gene|F76A03A71DADC32C7166E37B994EFED019FDF8A4|pyk]::kan trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKK29180 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CATTTGGTTCACTTCCTTCT, downstream forward: _UP4_TAATTACAGGTGAAAATGGA
Expression vectors
expression in ''E. coli'', N-terminal His-tag: pGP1100 (in [wiki|pWH844]), available in [wiki|Jörg Stülke]'s lab, [pubmed|20389117]
expression in ''B. subtilis'', native protein: pGP1411 (in [wiki|pBQ200]), available in [wiki|Jörg Stülke]'s lab
expression in ''B. subtilis'', N-terminal Strep-tag: pGP1409 (in [wiki|pGP380]), available in [wiki|Jörg Stülke]'s lab
expression in ''B. subtilis'', C-terminal Strep-tag: pGP1410 (in [wiki|pGP382]), available in [wiki|Jörg Stülke]'s lab
two-hybrid system
''B. pertussis'' adenylate cyclase-based bacterial two hybrid system ([wiki|BACTH]), available in [wiki|Jörg Stülke]'s lab, [pubmed|19193632]
lacZ fusion
see ''[gene|A7E31A5EBCF343B3A841E5F6E9CDE3CA5D1622EB|pfkA]
labs
[wiki|Jörg Stülke], University of Göttingen, Germany [http://wwwuser.gwdg.de/~genmibio/stuelke.html Homepage]
References
Page visits: 17092
Time of last update: 2024-12-21 14:28:17
Author of last update: Jstuelk