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Investigating the physiological roles and intracellular localization of an arginine kinase in Caenorhabditis elegans
Author(s) -
Thomas David H,
Snider Mark J
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a273-a
Subject(s) - arginine kinase , kinase , arginine , biochemistry , biology , polyclonal antibodies , gene isoform , microbiology and biotechnology , isozyme , subcellular localization , mitochondrion , caenorhabditis elegans , enzyme , amino acid , antibody , gene , genetics
Arginine kinase (AK) catalyzes the reversible phosphoryl transfer between ATP and arginine. It is the invertebrate homologue of creatine kinase, and is found primarily in cells that experience high and variable fluctuations in ATP hydrolysis. Five distinct isoforms of AK are recognized in the C. elegans proteome. A peptide multiple sequence alignment has identified one of these AKs as a likely mitochondrial arginine kinase, due to a high degree of similarity with that of established mitochondrial creatine kinases. If substantiated, the results presented here would represent the first biochemical evidence of a mitochondrial AK. The AK gene has been cloned, expressed in E. coli , and purified. Kinetic assays have shown the suspected enzyme to be a catalytically competent AK. A unique 10 amino acid segment of the AK was used to generate rabbit anti‐AK primary polyclonal antibodies. Western blots of the 5 recombinant C. elegans AK isozymes have proven antibody specificity. Immunohistochemical techniques using confocal microscopy are underway to determine the subcellular localization of this enzyme. This research is supported by NSF grant #0344432.