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Determining the subcellular localization of a novel arginine kinase in Caenorhabditis elegans
Author(s) -
Pipitone Baldassare D.,
Snider Mark J.
Publication year - 2006
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.20.5.a900-b
Subject(s) - arginine kinase , caenorhabditis elegans , biology , kinase , gene , microbiology and biotechnology , isozyme , polyclonal antibodies , biochemistry , mitochondrion , subcellular localization , arginine , enzyme , antibody , genetics , amino acid
Arginine kinase (AK) functions to catalyze the transfer of a high‐energy phosphoryl group between ATP and arginine. This process provides an ATP buffering capacity to cells with high energy demands. Five putative AK genes have been identified in the C. elegans genome, but have yet to be characterized. Peptide sequence alignments of these putative C. elegans AKs with known AKs and creatine kinases (the vertebrate homologue) suggest that one of the five enzymes in this nematode may be localized to the mitochondrion. The gene of this AK has been cloned and the recombinant enzyme expressed and purified from E. coli. Kinetic data indicate that the gene product is a catalytically competent arginine kinase. Immunohistochemical techniques will be conducted to provide evidence for mitochondrial localization of this AK within C. elegans . Peptide sequence alignments of the five C. elegans AKs predicted an antigenic sequence unique to only this AK isozyme. Rabbit anti‐AK polyclonal antibodies (Pacific Immunology) were generated and exhibited (by Western Blot) specific binding to the intended AK isozyme antigen. Future results of confocal fluorescence microscopy of nematodes treated with these antibodies may help to confirm mitochondrial localization of this enzyme, and further suggest the existence of a phosphoarginine shuttle in this organism. This research was supported by NSF grant #0344432.