title: Arginine kinase isoforms in the closest protozoan relative of metazoans.
abstract: The genome of the choanoflagellate Monosiga brevicollis contains at least three genes for the phosphoryl transfer enzyme, arginine kinase (AK; EC 2.7.3.3). Bioinformatic analyses of the deduced amino acid sequences of the proteins coded for by two of these genes showed that one of these AKs is cytoplasmic (denoted AK1) while the other appears to have an N-terminal mitochondrial targeting peptide (denoted AK2). Cloning and expression of the cDNA for AK1 yielded considerable soluble AK activity. Three AK2 constructs were expressed - one corresponding to the full length protein and two corresponding to truncated versions in which the signal peptide had been deleted. Expression of the former construct yielded minimal soluble activity. In contrast, significant AK activity was found in both truncated constructs confirming the importance of removal of the targeting peptide for proper folding and catalytic activity. Both AK1 and AK2 are functional oligomers unlike typical AKs which are monomeric. A phylogenetic analysis showed that these choanoflagellate AKs group more closely with a supercluster consisting of cytoplasmic and mitochondrial CKs and invertebrate AKs that evolved secondarily from a CK-like ancestor. Reaction-diffusion constraints in choanoflagellates are likely mitigated by the presence of AK isoforms which facilitate energy transport in these highly polarized cells.
authors: Institute of Molecular Biophysics, Department of Biological Science, Florida State University, Tallahassee, 32306, USA.
authors: Gregg G
authors: GG
authors: Hoffman
authors: W Ross
authors: WR
authors: Ellington
mesh_terms: D000595:Amino Acid Sequence; D001122:Arginine Kinase / Q000235:genetics* / Q000378:metabolism; D056897:Choanoflagellata / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D019143:Evolution, Molecular; D015870:Gene Expression; D008969:Molecular Sequence Data; D010802:Phylogeny; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D020033:Protein Isoforms; D015800:Protozoan Proteins; D001122:Arginine Kinase