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6.4.1.1: pyruvate carboxylase

This is an abbreviated version!
For detailed information about pyruvate carboxylase, go to the full flat file.

Word Map on EC 6.4.1.1

Reaction

ATP
+
pyruvate
 +
HCO3-
 +
H+
=
ADP
 +
phosphate
 +
oxaloacetate

Synonyms

Carboxylase, pyruvate, EhPYC1, HpPyc1p, Mfla_1512, MSmeg_2412, PC, Pcase, PCB, PCC, PCx, PYC, Pyc1, Pyc1p, PYC2, pycA, pyruvate carboxylase, pyruvate carboxylase 1, Pyruvic carboxylase, RePC

ECTree

     6 Ligases
         6.4 Forming carbon-carbon bonds
             6.4.1 Ligases that form carbon-carbon bonds (only sub-subclass identified to date)
                6.4.1.1 pyruvate carboxylase

Cloned

Cloned on EC 6.4.1.1 - pyruvate carboxylase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
a Pseudomonas aeruginosa strain carrying the T7 polymerase gene can serve as a host for the overexpression of Mycobacterium smegmatis alpha4 under the control of the T7 promoter from a broad-host-range conjugative plasmid. Overexpression occurrs both in aerobic (LB medium) and nitrate-respiring anaerobic (LB medium plus glucose and nitrate) cultures. The latter system presents a simpler option because it involved room temperature cultures in stationary screw-cap bottles. Developed of a Pseudomonas aeruginosa DELTApyc strain that allows the expression of recombinant PYCs in the absence of the native enzyme
-
a Pseudomonas aeruginosa strain carrying the T7 polymerase gene can serve as a host for the overexpression of Pseudomonas aeruginosa alpha4beta4 PYC under the control of the T7 promoter from a broad-host-range conjugative plasmid. Overexpression occurs both in aerobic (LB medium) and nitrate-respiring anaerobic (LB medium plus glucose and nitrate) cultures. The latter system presents a simpler option because it involved room temperature cultures in stationary screw-cap bottles. Development of a Pseudomonas aeruginosa DELTApyc strain that allows the expression of recombinant PYCs in the absence of the native enzyme
-
codon-optimized expression in CHO cells
-
coexpressed with human erythropoietin in BHK-21 cells
-
coexpression of pyruvate carboxylase 1 isozyme (Pyc1) with an N-terminal myc tag, together with constructs encoding either the biotin carboxylase domain or the transcarboxylase-biotin carboxyl carrier domain, each with an N-terminal 9-histidine tag
DNA and amino acid sequence determination and analysis
DNA and amino acid sequence determination and analysis, expression mutant enzymes and of the isolated biotin carboxylase domain
-
DNA and amino acid sequence determination and analysis, genetic structure, key cognate transcription factors regulating tissue-specific expression, transcriptional regulation, overview
DNA and amino acid sequence determination and analysis, genetic structure, key cognate transcription factors regulating tissue-specific expression. Five species of enzyme mRNAs have been reported, each having the same coding sequence but differing in their 5'-untranslated regions. These mRNA variants are the product of alternative splicing of two primary transcripts initiated from two alternative promoters, the proximal and the distal promoters. Neither of these promoters contains a TATA box but both possess multiple GC boxes. Production of specific forms of PC mRNA are linked to certain physiological states, i.e. development, gluconeogenesis and lipogenesis. Two pancreatic isletspecific transcription factors, i.e. pancreatic duodenal homeobox-1or PDX1, and v-MAFA, are involved in transcriptional regulation of the enzyme in INS1 cells. Identification of a putative cAMP-responsive element in the proximal promoter of the rat PC gene, transcriptional regulation, overview
-
DNA and amino acid sequence determination and analysis, identification of promoter regions of bovine pyruvate carboxylase, putative transcription factor binding sites of promoter P1 and of P2, overview
expressed in 293T cells
expressed in Escherichia coli
expressed in Escherichia coli and Streptomyces lividans as a His-tagged fusion protein. In Streptomyces lividans a functional His-EhPYC1 is expressed
expressed in Escherichia coli as a His-tagged fusion protein
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli NZN111 and AFP111
-
expressed in L-929 cells
-
expression analysis of the enzyme in diverse wild-type and knockout insulinoma cell lines, overview
expression in CHO-K1-hGM-CSF cells
-
expression in Escherichia coli GJT001, coexpression with pantothenate kinase
-
expression in Escherichia coli JM109
expression in Escherichia coli mutant SBS110MG
-
expression in Escherichia coli ptsG mutant
-
expression in Escherichia coli wild-type GJT001 and mutants YBS121
-
expression in type B pyruvate decarboxylase deficient skin fibroblasts
-
expression of His9-tagged wild-type and mutant enzymes in Escherichia coli strain BL21
-
expression of the C-terminal region, excluding the mitochondrial targeting sequence, in Escherichia coli strain BL21(DE3)
expression of the subunits biotin carboxylase BC and carboxyl transferase CT (PC-(BC+CT)) and biotin carboxyl carrier protein BCCP (PC-(BCCP)) in Escherichia coli JM109
-
FLAG-tagged human pyruvate carboxylase is introduced into a dihydrofolate-deficient CHO cell line DG44. Through the expression of the human pyruvate carboxylase enzyme, lactate formation in CHO cell culture can be efficiently reduced. This effect of expression of the human pyruvate carboxylase is observed not only in adherent batch culture using the serum-containing medium but in the serum-free suspension fed-batch culture as well, demonstrating its potential use to extend the culture longevity of CHO cell culture, which often shows a significant accumulation of lactate
genes pycA and pycB encoding two subunits are divergently transcribed upstream of pycR encoding for a pyruvate carboxylase regulator PycR, genomic organization of the PA5436-5435, pycAB, operon and the PA5437 or pycR gene, overview
-
genetic structure, expression analysis, genotyping of genetic variants
-
key cognate transcription factors regulating tissue-specific expression. The proximal promoter of the bovine PC gene mediates the mRNA variants that are restricted to gluconeogenic and lipogenic tissues, transcriptional regulation, overview
-
Pyc1 isoform and C249A mutant
-
stable overexpression in enzyme-deficient INS-1 insulinoma cell line
-
the enzyme is expressed in 6 alternatively spliced variants that share a common ORF but differ in the 5'UTRs, resulting in different translational efficiencies, overview. In vitro transcription and translation by rabbit reticulocyte lysate as luciferase-linked protein, functional synthesis of luciferase, overview
-
two genes PYC1 and PYC2 located on different chromosomes, expression of PYC1 and PYC2 is influenced by both the growth phase and carbon source, overview
-
two polypeptide chain: PC-(BC) and PC-(CT+BCCP)
-
wild-type and mutants without enzyme activity
-