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Protein

Phosphoenolpyruvate carboxykinase [GTP]

Gene

pckG

Organism
Mycobacterium smegmatis
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Involved in the gluconeogenesis, in growth on fatty acids and is important for initiation of infection in the macrophages. Catalyzes the GTP-dependent conversion of oxaloacetate (OAA) to phosphoenolpyruvate (PEP), the rate-limiting step in the metabolic pathway that produces glucose from lactate and other precursors derived from the citric acid cycle. It can also use IDP or ADP, but with a lower activity.2 Publications

Catalytic activityi

GTP + oxaloacetate = GDP + phosphoenolpyruvate + CO2.2 Publications

Cofactori

Mn2+2 PublicationsNote: Binds 1 Mn(2+) ion per subunit. Can also use Co2+ ion.2 Publications

Enzyme regulationi

Inhibited by oxalate and by alpha-ketoglutarate. In vitro, the enzyme is stimulated by reducing agents such as dithiothreitol (DTT), reduced glutathione and 2-mercaptoethanol.1 Publication

Kineticsi

  1. KM=0.67 µM for OAA (PEP forming activity at pH 7.2 and 37 degrees Celsius).1 Publication
  2. KM=3.3 µM for manganese (OAA forming activity at pH 7.2 and 37 degrees Celsius).1 Publication
  3. KM=69 µM for GDP (OAA forming activity at pH 7.2 and 37 degrees Celsius).1 Publication
  4. KM=132 µM for manganese (OAA forming activity without DTT at pH 7.2 and 37 degrees Celsius).1 Publication
  5. KM=208 µM for cobalt (OAA forming activity without DTT at pH 7.2 and 37 degrees Celsius).1 Publication
  6. KM=471 µM for PEP (OAA forming activity at pH 7.2 and 37 degrees Celsius).1 Publication
  1. Vmax=8.6 µmol/min/mg enzyme with manganese as substrate (OAA forming activity without DTT at pH 7.2 and 37 degrees Celsius).1 Publication
  2. Vmax=9.5 µmol/min/mg enzyme with cobalt as substrate (OAA forming activity without DTT at pH 7.2 and 37 degrees Celsius).1 Publication
  3. Vmax=21 µmol/min/mg enzyme with OAA as substrate (PEP forming activity at pH 7.2 and 37 degrees Celsius).1 Publication
  4. Vmax=32 µmol/min/mg enzyme with manganese as substrate (OAA forming activity at pH 7.2 and 37 degrees Celsius).1 Publication
  5. Vmax=33.6 µmol/min/mg enzyme with GDP as substrate (OAA forming activity at pH 7.2 and 37 degrees Celsius).1 Publication
  6. Vmax=38.2 µmol/min/mg enzyme with PEP as substrate (OAA forming activity at pH 7.2 and 37 degrees Celsius).1 Publication

pH dependencei

Optimum pH is between 7 and 7.4.1 Publication

Temperature dependencei

Optimum temperature is 70 degrees Celsius.1 Publication

Pathway:igluconeogenesis

This protein is involved in the pathway gluconeogenesis, which is part of Carbohydrate biosynthesis.Curated
View all proteins of this organism that are known to be involved in the pathway gluconeogenesis and in Carbohydrate biosynthesis.

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Binding sitei81 – 811SubstrateUniRule annotation
Metal bindingi229 – 2291ManganeseUniRule annotation
Metal bindingi249 – 2491Manganese; via tele nitrogenUniRule annotation
Binding sitei271 – 2711SubstrateUniRule annotation
Active sitei273 – 2731UniRule annotation
Metal bindingi296 – 2961ManganeseUniRule annotation
Binding sitei388 – 3881GTPUniRule annotation
Binding sitei419 – 4191GTPUniRule annotation

Regions

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Nucleotide bindingi272 – 2776GTPUniRule annotation
Nucleotide bindingi514 – 5174GTPUniRule annotation

GO - Molecular functioni

GO - Biological processi

Complete GO annotation...

Keywords - Molecular functioni

Decarboxylase, Lyase

Keywords - Biological processi

Gluconeogenesis

Keywords - Ligandi

GTP-binding, Manganese, Metal-binding, Nucleotide-binding

Enzyme and pathway databases

BRENDAi4.1.1.32. 3512.
SABIO-RKQ9AGJ6.
UniPathwayiUPA00138.

Names & Taxonomyi

Protein namesi
Recommended name:
Phosphoenolpyruvate carboxykinase [GTP]1 PublicationUniRule annotation (EC:4.1.1.32UniRule annotation2 Publications)
Short name:
PEP carboxykinase1 PublicationUniRule annotation
Short name:
PEPCK1 PublicationUniRule annotation
Alternative name(s):
GTP-dependent phosphoenolpyruvate carboxykinase1 Publication
Short name:
GTP-PEPCK1 Publication
Gene namesi
Name:pckG
Synonyms:pck
OrganismiMycobacterium smegmatis
Taxonomic identifieri1772 [NCBI]
Taxonomic lineageiBacteriaActinobacteriaCorynebacterialesMycobacteriaceaeMycobacterium

Subcellular locationi

GO - Cellular componenti

Complete GO annotation...

Keywords - Cellular componenti

Cytoplasm

Pathology & Biotechi

Disruption phenotypei

Cells lacking this gene are inhibited when they grow on acetate and palmitate.1 Publication

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi75 – 751D → A: Significantly reduced carboxykinase activity. The affinity for Mn(2+), OAA and PEP decreases 9-, 2- and 3-fold compared to the wild-type, respectively. 1 Publication
Mutagenesisi75 – 751D → N: Significantly reduced carboxykinase activity. The affinity for Mn(2+), OAA and PEP decreases 3.5-, 2.8- and 3-fold compared to the wild-type, respectively. 1 Publication
Mutagenesisi75 – 751D → Q: Significantly reduced carboxykinase activity. The affinity for PEP and OAA decreases 9- and 2-fold compared to the wild-type, respectively. 1 Publication
Mutagenesisi75 – 751D → S: Significantly reduced carboxykinase activity. The affinity for Mn(2+), OAA and PEP decreases 3-, 2- and 3-fold compared to the wild-type, respectively. 1 Publication
Mutagenesisi78 – 781D → A: Exhibits a very low activity in either the OAA- or the PEP-forming direction. With Mn(2+) at 0.2 mM, the specific OAA-forming activity is 0.3% that of the wild-type. This value increases to 0.5%, when the Mn(2+) concentration raises to 2 mM. With Mn(2+) at 0.2 mM, the specific PEP-forming activity is 0.4% that of the wild-type. 1 Publication
Mutagenesisi83 – 831E → A: Exhibits a very low activity in either the OAA- or the PEP-forming direction. With Mn(2+) at 0.2 mM, the specific OAA-forming activity is 0.9% that of the wild-type. This value increases to 3.4%, when the Mn(2+) concentration raises to 2 mM. With Mn(2+) at 0.2 mM, the specific PEP-forming activity is 2.3% that of the wild-type. The affinity for PEP is not significantly different from that of the wild-type, but the affinity for OAA and GDP decreases 2-fold. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 605605Phosphoenolpyruvate carboxykinase [GTP]PRO_0000103610Add
BLAST

Interactioni

Subunit structurei

Monomer.1 Publication

Protein-protein interaction databases

STRINGi246196.MSMEG_0255.

Structurei

3D structure databases

ProteinModelPortaliQ9AGJ6.
SMRiQ9AGJ6. Positions 12-604.
ModBaseiSearch...
MobiDBiSearch...

Family & Domainsi

Region

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Regioni220 – 2223Substrate bindingUniRule annotation
Regioni386 – 3883Substrate bindingUniRule annotation

Sequence similaritiesi

Family and domain databases

Gene3Di3.40.449.10. 1 hit.
3.90.228.20. 2 hits.
HAMAPiMF_00452. PEPCK_GTP.
InterProiIPR018091. PEP_carboxykin_GTP_CS.
IPR013035. PEP_carboxykinase_C.
IPR008209. PEP_carboxykinase_GTP.
IPR008210. PEP_carboxykinase_N.
[Graphical view]
PANTHERiPTHR11561. PTHR11561. 1 hit.
PfamiPF00821. PEPCK. 1 hit.
[Graphical view]
PIRSFiPIRSF001348. PEP_carboxykinase_GTP. 1 hit.
SUPFAMiSSF68923. SSF68923. 1 hit.
PROSITEiPS00505. PEPCK_GTP. 1 hit.
[Graphical view]

Sequencei

Sequence statusi: Complete.

Q9AGJ6-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MTSATIPGLD TAPTKHQGLL AWVQEVAELT QPDRVVFADG SDEEYERLCA
60 70 80 90 100
HLVEAGTFQK LNPEKQPNSY LALSDPSDVA RVESRTFICT EREIDAGPTN
110 120 130 140 150
NWMDPAEMRG IMTDLYRGSM RGRTLYVVPF CMGPLDAEDP KLGVEITDSE
160 170 180 190 200
YVVVSMRTMT RMGRAALDKL GDDGFFVKAL HSIGAPLEPG QKDVPWPCND
210 220 230 240 250
TKYITHFPET REIWSFGSGY GGNALLGKKC YSLRIASAMA HDEGWLAEHM
260 270 280 290 300
LILKLISPEN KAYFIAAAFP SACGKTNLAM LQPTIEGWRA ETVGDDIAWM
310 320 330 340 350
RFGKDGRLYA TNPEFGFFGV APGTNWSSNP NAMKTIAAGN TVFTNVAKTD
360 370 380 390 400
DGDVWWEGLE GDPQHLIDWK GNDWTPESGE KAAHPNSRYC TPISQCPTLA
410 420 430 440 450
PEWDDPQGVP ISAILFGGRR KTTVPLITEA RDWQHGVFIG ATLGSEQTAA
460 470 480 490 500
AEGKVGTVRR DPMAMLPFLG YNVGDYFAHW INVGKNADES KLPKVFFVNW
510 520 530 540 550
FRRGDDGRFL WPGFGENSRV LKWAVERIEH KADGKSTPIG IVPTAADLDL
560 570 580 590 600
EGLDVDPADV DEALAVKPEE WRAELPLIEE WFEFVGEKLP TGLKDEFDAL

KERLG
Length:605
Mass (Da):66,944
Last modified:June 1, 2001 - v1
Checksum:iE07A46D4FE35EDC0
GO

Mass spectrometryi

Molecular mass is 71209 Da from positions 1 - 605. Determined by MALDI. 1 Publication

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AF332191 Genomic DNA. Translation: AAK28534.1.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AF332191 Genomic DNA. Translation: AAK28534.1.

3D structure databases

ProteinModelPortaliQ9AGJ6.
SMRiQ9AGJ6. Positions 12-604.
ModBaseiSearch...
MobiDBiSearch...

Protein-protein interaction databases

STRINGi246196.MSMEG_0255.

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Enzyme and pathway databases

UniPathwayiUPA00138.
BRENDAi4.1.1.32. 3512.
SABIO-RKQ9AGJ6.

Family and domain databases

Gene3Di3.40.449.10. 1 hit.
3.90.228.20. 2 hits.
HAMAPiMF_00452. PEPCK_GTP.
InterProiIPR018091. PEP_carboxykin_GTP_CS.
IPR013035. PEP_carboxykinase_C.
IPR008209. PEP_carboxykinase_GTP.
IPR008210. PEP_carboxykinase_N.
[Graphical view]
PANTHERiPTHR11561. PTHR11561. 1 hit.
PfamiPF00821. PEPCK. 1 hit.
[Graphical view]
PIRSFiPIRSF001348. PEP_carboxykinase_GTP. 1 hit.
SUPFAMiSSF68923. SSF68923. 1 hit.
PROSITEiPS00505. PEPCK_GTP. 1 hit.
[Graphical view]
ProtoNetiSearch...

Publicationsi

  1. "A GTP-dependent vertebrate-type phosphoenolpyruvate carboxykinase from Mycobacterium smegmatis."
    Mukhopadhyay B., Concar E.M., Wolfe R.S.
    J. Biol. Chem. 276:16137-16145(2001) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-13, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION, COFACTOR, MASS SPECTROMETRY, SUBUNIT, SUBSTRATE SPECIFICITY.
  2. "pckA-deficient Mycobacterium bovis BCG shows attenuated virulence in mice and in macrophages."
    Liu K., Yu J., Russell D.G.
    Microbiology 149:1829-1835(2003) [PubMed] [Europe PMC] [Abstract]
    Cited for: DISRUPTION PHENOTYPE.
  3. "Roles of Asp75, Asp78, and Glu83 of GTP-dependent phosphoenolpyruvate carboxykinase from Mycobacterium smegmatis."
    Case C.L., Concar E.M., Boswell K.L., Mukhopadhyay B.
    J. Biol. Chem. 281:39262-39272(2006) [PubMed] [Europe PMC] [Abstract]
    Cited for: FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ASP-75; ADP-78 AND GLU-83, COFACTOR.

Entry informationi

Entry nameiPCKG_MYCSM
AccessioniPrimary (citable) accession number: Q9AGJ6
Entry historyi
Integrated into UniProtKB/Swiss-Prot: September 26, 2001
Last sequence update: June 1, 2001
Last modified: July 22, 2015
This is version 69 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Keywords - Technical termi

Direct protein sequencing

Documents

  1. PATHWAY comments
    Index of metabolic and biosynthesis pathways
  2. SIMILARITY comments
    Index of protein domains and families

External Data

Dasty 3

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.