pycA
168
pyruvate carboxylase
locus
BSU_14860
Molecular weight
127.72 kDa
pI
5.41
function
replenishment of the oxaloacetate pool
product
pyruvate carboxylase
essential
no
synonyms
pycA, ylaP
Outlinks
Genomic Context
Categories containing this gene/protein
List of homologs in different organisms, belongs to COG1038 (Galperin et al., 2021)
This gene is a member of the following regulons
Gene
Coordinates
1,554,185 1,557,631
The protein
Catalyzed reaction/ biological activity
ATP + hydrogencarbonate + pyruvate --> ADP + H+ + oxaloacetate + phosphate (according to UniProt)
[wiki|Domains]
[wiki|ATP-grasp domain] (aa 125-321) (according to UniProt)
Biotin carboxylation domain (aa 1-457) (according to UniProt)
Pyruvate carboxyltransferase domain (aa 534-802) (according to UniProt)
Biotinyl-binding domain (aa 1071-1146) (according to UniProt)
[wiki|Cofactors]
biotin [Pubmed|24816803]
Structure
[PDB|3BG5] (''S. aureus'') [pubmed|18297087]
[AF|Q9KWU4]
Effectors of protein activity
allosteric activation by acetyl-CoA [pubmed|22120519,4146915]
feedback inhibition by aspartate [pubmed|35130724]
Apo-[protein|90AFF93D6E7BC993B2773C12EE400C80A87A4831|darB] stimulates [protein|6ABF79C6EEF078E4F71F569FACCA80DA67191AE2|pycA] activity (by increasing Vmax of the reaction) [pubmed|35130724]
Kinetic information
Km for pyruvate (in the presence of acetyl-CoA): 0.23 mM [pubmed|35130724]
Vmax 42.7 U/ mg of protein (in the absence of [protein|90AFF93D6E7BC993B2773C12EE400C80A87A4831|darB]) [pubmed|35130724]
Vmax 87.9 U/ mg of protein (in the presence of [protein|90AFF93D6E7BC993B2773C12EE400C80A87A4831|darB]) [pubmed|35130724]
[wiki|Interactions]
interaction with apo-[protein|90AFF93D6E7BC993B2773C12EE400C80A87A4831|darB] under conditions of potassium starvation activates [protein|6ABF79C6EEF078E4F71F569FACCA80DA67191AE2|pycA] [pubmed|35130724]
the C-terminal part of [protein|6ABF79C6EEF078E4F71F569FACCA80DA67191AE2|pycA] is required for the interaction with [protein|90AFF93D6E7BC993B2773C12EE400C80A87A4831|darB] [pubmed|35130724]
[wiki|Localization]
membrane associated [Pubmed|18763711]
cytoplasm (homogeneously distributed throughout the cell) [Pubmed|24825009]
Expression and Regulation
Operons
genes
[gene|8AB7D225DBEE6BD695A4A8A2384D2C029B571C57|ftsW]-[gene|6ABF79C6EEF078E4F71F569FACCA80DA67191AE2|pycA]
description
[pubmed|22383849]
regulation
constitutively expressed [Pubmed|23701187]
regulatory mechanism
[protein|6FDA046DCBBAE4588C4BC7163A04B9E6AEDBAC55|yofA]: activation, [Pubmed|17526699], in [regulon|protein:6FDA046DCBBAE4588C4BC7163A04B9E6AEDBAC55|yofA regulon]
stringent response: positive regulation, due to presence of adenines at +1 and +2 positions of the transcript [Pubmed|20081037], in [regulon|other_regulator:stringent response|stringent response]
[protein|857EF1AC54DEA0690C01B7F8CEFFD16312C58F20|tnrA]: repression, in [regulon|protein:857EF1AC54DEA0690C01B7F8CEFFD16312C58F20|tnrA regulon]
sigma factors
[protein|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|sigA]: sigma factor, [Pubmed|20081037], in [regulon|protein:360F48D576DE950DF79C1A2677B7A35A8D8CC30C|sigA regulon]
genes
[gene|6ABF79C6EEF078E4F71F569FACCA80DA67191AE2|pycA]
description
[Pubmed|20081037]
regulation
subject to positive stringent control upon lysine starvation [Pubmed|20081037]
regulatory mechanism
[protein|A8C82F5825DE0921DA6B4ACDE0CC31EC7749273C|purR]: repression, [pubmed|34967415], in [regulon|protein:A8C82F5825DE0921DA6B4ACDE0CC31EC7749273C|purR regulon]
Biological materials
Mutant
MGNA-A544 (ylaP::erm), available at the [https://shigen.nig.ac.jp/bsub/resource/strainGeneDisrupted/detail/544 NBRP B. subtilis, Japan]
GP793 (cat), available in [wiki|Jörg Stülke]'s lab
BKE14860 ([gene|6ABF79C6EEF078E4F71F569FACCA80DA67191AE2|pycA]::erm trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKE14860 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CAAACTTTTCTCCCCCTTAC, downstream forward: _UP4_TAAAAAACAAAGAGTGTATA
BKK14860 ([gene|6ABF79C6EEF078E4F71F569FACCA80DA67191AE2|pycA]::kan trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKK14860 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CAAACTTTTCTCCCCCTTAC, downstream forward: _UP4_TAAAAAACAAAGAGTGTATA
Expression vectors
pGP1289 (N-terminal Strep-tag, purification from ''B. subtilis'', for [wiki|SPINE], in [wiki|pGP380]), available in [wiki|Jörg Stülke]'s lab
pGP3322 (no tag, purification from B. subtilis, in [wiki|pGP382]) , available in [wiki|Jörg Stülke]'s lab
pGP3458 (expression in E. coli, in pET28a), available in [wiki|Jörg Stülke]'s lab
two-hybrid system
available in [wiki|Jörg Stülke]'s lab
lacZ fusion
pGP3328 (cat, based on [wiki|pAC5]]), available in [wiki|Jörg Stülke]'s lab
References
Reviews
Original Publications
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Time of last update: 2024-12-21 00:18:52
Author of last update: Jstuelk