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PMID:10930737

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Citation

Guo, K and Newell, PC (2000) Pyridoxal kinase knockout of Dictyostelium complemented by the human homologue. FEMS Microbiol. Lett. 189:195-200

Abstract

The gene (pykA) encoding pyridoxal kinase which converts pyridoxal (vitamin B(6)) to pyridoxal phosphate was isolated from Dictyostelium discoideum using insertional mutagenesis. Cells of a pykA gene knockout grew poorly in axenic medium with low yield but growth was restored by the addition of pyridoxal phosphate. Sequencing indicated a gene, with one intron, encoding a predicted protein of 301 amino acids that was 42% identical in amino acid sequence to human pyridoxal kinase. After expression of the wild-type gene in Escherichia coli, the purified PykA protein product was shown to have pyridoxal kinase enzymatic activity with a K(m) of 8.7 microM for pyridoxal. Transformation of the Dictyostelium knockout mutant with the human pyridoxal kinase gene gave almost the same level of complementation as that seen using transformation with the wild-type Dictyostelium gene. Phylogenetic analysis indicated that the Dictyostelium amino acid sequence was closer to human pyridoxal kinase than to pyridoxal kinases of lower eukaryotes.

Links

PubMed

Keywords

Amino Acid Sequence; Animals; Dictyostelium/genetics; Gene Expression Regulation, Enzymologic; Humans; Molecular Sequence Data; Mutagenesis, Insertional; Pyridoxal Kinase/genetics; Sequence Alignment; Sequence Homology, Amino Acid

Significance

Annotations

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

DICDI:PDXK

acts_upstream_of_or_within

GO:0019954: asexual reproduction

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:PDXK

acts_upstream_of_or_within

GO:0031152: aggregation involved in sorocarp development

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:PDXK

enables

GO:0008478: pyridoxal kinase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DICDI:PDXK

enables

GO:0008478: pyridoxal kinase activity

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:O00764

F

Seeded From UniProt

complete

DICDI:PDXK

acts_upstream_of_or_within

GO:0042816: vitamin B6 metabolic process

ECO:0000305: curator inference used in manual assertion

GO:0008478

P

Seeded From UniProt

complete


See also

References

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