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PMID:2110163

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Citation

Weldon, SL, Rando, A, Matathias, AS, Hod, Y, Kalonick, PA, Savon, S, Cook, JS and Hanson, RW (1990) Mitochondrial phosphoenolpyruvate carboxykinase from the chicken. Comparison of the cDNA and protein sequences with the cytosolic isozyme. J. Biol. Chem. 265:7308-17

Abstract

The amino acid sequence of the mitochondrial form of phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK-M) from the chicken was deduced from the 3571 nucleotide sequence of three overlapping cDNA clones. The derived protein sequence, which includes 607 amino acids of the mature enzyme and a leader sequence, was aligned with nine tryptic peptides of PEPCK-M and the primary sequence of the cytosolic form of PEPCK from chicken. Secondary structure predictions for the two PEPCK isozymes indicated similar packing elements of conserved, hydrophobic beta strands in the central core of the primary sequence. This core protein, which contained three GTP-binding consensus elements, was 80% identical in the two chicken isozymes, although the overall level of identity was only 63% for amino acids and 60% for nucleotides. The untranslated regions of the two cDNAs were dissimilar, although both mRNAs have potential for significant secondary structure. The PEPCK-M mRNA contained several G-C-rich regions which demonstrated free energies of formation in dyad symmetry programs up to -70 kcal/mol. The 1.6-kilobase (kb) 3'-untranslated region contained several repeat elements including one of 11 base pairs, which was present 30 times; but, a signal sequence for polyadenylation was not present. Each of the three PEPCK-M cDNA clones recognized two mRNAs of 4.2 and 3.4 kb in the livers and kidneys of starved or normally fed chickens. However, the level of these two related PEPCK-M mRNAs changed in response to cAMP treatment, with the larger mRNA predominant at 20 and 160 min and the 3.4-kb mRNA present at intermediate times. In contrast, the level of the 2.8-kb PEPCK-C mRNA increased dramatically upon addition of the cyclic nucleotide, particularly in the liver where it was not detected without cAMP induction. Thus, PEPCK-M and PEPCK-C, clearly represented the products of two distinct genes, which were distinguished by altered protein sequences and non-cross-hybridizing, differentially regulated mRNAs.

Links

PubMed

Keywords

Amino Acid Sequence; Animals; Base Sequence; Blotting, Northern; Chickens; Cytosol/enzymology; DNA/genetics; GTP-Binding Proteins/metabolism; Gene Library; Isoenzymes/genetics; Isoenzymes/metabolism; Liver/enzymology; Mitochondria, Liver/enzymology; Molecular Sequence Data; Oligonucleotide Probes; Peptide Fragments/isolation & purification; Phosphoenolpyruvate Carboxykinase (GTP)/genetics; Phosphoenolpyruvate Carboxykinase (GTP)/metabolism; RNA, Messenger/genetics; Restriction Mapping; Sequence Homology, Nucleic Acid

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

CHICK:PCKGC

enables

GO:0030145: manganese ion binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGC

enables

GO:0047134: protein-disulfide reductase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGC

enables

GO:0008906: inosine kinase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGC

involved_in

GO:0046015: regulation of transcription by glucose

ECO:0000304: author statement supported by traceable reference used in manual assertion


P

Seeded From UniProt

complete

CHICK:PCKGC

involved_in

GO:0051591: response to cAMP

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

CHICK:PCKGC

involved_in

GO:0015701: bicarbonate transport

ECO:0000245: automatically integrated combinatorial evidence used in manual assertion

P

Seeded From UniProt

complete

CHICK:PCKGC

involved_in

GO:0043950: positive regulation of cAMP-mediated signaling

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

CHICK:PCKGC

enables

GO:0019904: protein domain specific binding

ECO:0000245: automatically integrated combinatorial evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGC

enables

GO:0005525: GTP binding

ECO:0000245: automatically integrated combinatorial evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGM

enables

GO:0008906: inosine kinase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGM

involved_in

GO:0043687: post-translational protein modification

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

CHICK:PCKGM

enables

GO:0047134: protein-disulfide reductase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGM

enables

GO:0005525: GTP binding

ECO:0000245: automatically integrated combinatorial evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGM

involved_in

GO:0015701: bicarbonate transport

ECO:0000245: automatically integrated combinatorial evidence used in manual assertion

P

Seeded From UniProt

complete

CHICK:PCKGM

involved_in

GO:0043950: positive regulation of cAMP-mediated signaling

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

CHICK:PCKGM

enables

GO:0030145: manganese ion binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGM

enables

GO:0003729: mRNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGM

enables

GO:0004457: lactate dehydrogenase activity

ECO:0000303: author statement without traceable support used in manual assertion

F

Seeded From UniProt

complete

CHICK:PCKGM

involved_in

GO:0051591: response to cAMP

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

CHICK:PCKGM

involved_in

GO:0030150: protein import into mitochondrial matrix

ECO:0000245: automatically integrated combinatorial evidence used in manual assertion

P

Seeded From UniProt

complete

CHICK:PCKGM

enables

GO:0019904: protein domain specific binding

ECO:0000245: automatically integrated combinatorial evidence used in manual assertion

F

Seeded From UniProt

complete

Notes

See also

References

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