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Of the six phosphagen kinases found in animals, the primary structure is known only for creatine kinase. Here we report three cDNA-derived or chemically determined amino acid sequences of two kinds of phosphagen kinases: a glycocyamine kinase from the polychaete Neanthes diversicolor (Annelida) and arginine kinase from the abalone Nordotis madaka (Mollusca) and the shrimp Penaeus japonicus (Arthropoda). Like vertebrate creatine kinases are monomers. These enzymes consist of 350 to 390 amino acid residues, and have a calculated molecular mass of 39,900 to 44,500 Da. Neanthes glycocyamine kinase shows 50 to 58% sequence similarity with vertebrate and invertebrate creatine kinases, having the greatest similarity (57 to 58%) with vertebrate mitochondrial creatine kinase isoform. It shows lower, but significant similarity (37 to 39%) with invertebrate arginine kinases. The sequence similarity between Nordotis and Penaeus arginine kinases is 51%. A phylogenetic tree constructed from 14 amino acid sequences of phosphagen kinases showed that they can be separated into three major clusters corresponding to creatine kinase, glycocyamine kinase and arginine kinase. The cluster of glycocyamine kinase is apparently closer to that of creatine kinase than arginine kinase. The cluster of creatine kinase is composed of several subclusters, each corresponding to three vertebrate isoforms and the invertebrate enzyme.
J Mol Biol 1994 Apr 01
PMID:Evolution of phosphagen kinase. Primary structure of glycocyamine kinase and arginine kinase from invertebrates. 814 48

Dimeric rabbit muscle creatine kinase (MM-CK) was bound to CNBr-activated Sepharose 4B by one of its subunits (MM-CKA). Treatment of MM-CKA with guanidine hydrochloride released the unbound subunit to yield the matrix-bound monomer (M-CKB). M-CKB recombined with dissociated MM-CK soluble subunits to reconstitute a matrix-bound dimer (MM-CKC). M-CKB also associated with dissociated subunits of BB-CK from crude extracts of rabbit brain and of arginine kinase from sea cucumber muscle (MM-AK) to form the matrix-bound heterohybrids MB-CKC and M-CK/M-AKC, respectively. Guanidine hydrochloride gradient elution studies showed that MM-CKA, MM-CKC and MB-CKC were all dissociated at the same concentration of the denaturant (0.96 M), while the M-CK/M-AKC heterohybrid was less stable, dissociating at 0.5 M. The specific interaction between subunits of echinoderm and mammalian phosphagen kinases to form a hybrid enzyme of dual substrate specificity supports the view that these enzymes had a common evolutionary origin.
Comp Biochem Physiol Biochem Mol Biol 1994 May
PMID:Hybridization of matrix-bound MM-creatine kinase with BB-creatine kinase and arginine kinase. 820 93

Creatine kinase (CK) is coded for by at least four loci in higher vertebrates--two cytoplasmic isoforms, muscle (M) and brain (B), and two mitochondrial isoforms, sarcomeric and ubiquitous. M is expressed primarily in skeletal muscle, while B is expressed in a variety of cells, including cardiac and smooth muscle fibers, neurons, transport epithelia, and photoreceptors. M and B subunits form very stable homodimers (MM [M-CK], BB [B-CK]) and heterodimers (MB). M-CK is capable of binding to the M line of the myofibril, thereby creating an energy transfer microcompartment; BB and MB CKs are not. M- and B-like CKs are present in all vertebrates yet examined, including fish. Cytoplasmic, dimeric CKs are widely distributed in the invertebrates. The only available amino acid sequence for an invertebrate dimeric CK, that of the protostome polychaete Chaetopterus variopedatus, is just as similar to the vertebrate M isoform as to the B isoform. Echinoderms lack dimeric, cytoplasmic CKs, which appear to be replaced by a dimeric arginine kinase which evolved secondarily from CK. Thus, it is likely that the gene duplication event producing the M and B isoforms occurred after the divergence of the chordates from echinoderms. To narrow down the timing of this duplication event, we obtained the cDNA and deduced amino acid sequences of dimeric CKs from the tunicate Ciona intestinalis (subphylum Urochordata) and the lancelet Branchiostoma floridae (subphylum Cephalochordata). Our results show that these CKs are strikingly similar to both invertebrate and vertebrate CKs. However, phylogenetic analyses by neighbor-joining and parsimony show that these two enzymes appeared to have diverged before the point of divergence of the M and B isoforms. Thus, the gene duplication event for formation of the muscle and brain isoforms of CK most likely occurred during the radiation of the fish, a time noted for gene duplication events at a variety of other loci.
Mol Biol Evol 2001 Jul
PMID:Gene duplication events producing muscle (M) and brain (B) isoforms of cytoplasmic creatine kinase: cDNA and deduced amino acid sequences from two lower chordates. 1142 Mar 69

To test whether gaps resulting from sequence alignment contain phylogenetic signal concordant with those of base substitutions, we analyzed the occurrence of indel mutations upon a well-resolved, substitution-based tree for three nuclear genes in bumble bees (Bombus, Apidae: Bombini). The regions analyzed were exon and intron sequences of long-wavelength rhodopsin (LW Rh), arginine kinase (ArgK), and elongation factor-1alpha (EF-1alpha) F2 copy genes. LW Rh intron had only a few uninformative gaps, ArgK intron had relatively long gaps that were easily aligned, and EF-1alpha intron had many short gaps, resulting in multiple optimal alignments. The unambiguously aligned gaps within ArgK intron sequences showed no homoplasy upon the substitution-based tree, and phylogenetic signals within ambiguously aligned regions of EF-1alpha intron were highly congruent with those of base substitutions. We further analyzed the contribution of gap characters to phylogenetic reconstruction by incorporating them in parsimony analysis. Inclusion of gap characters consistently improved support for nodes recovered by substitutions, and inclusion of ambiguously aligned regions of EF-1alpha intron resolved several additional nodes, most of which were apical on the phylogeny. We conclude that gaps are an exceptionally reliable source of phylogenetic information that can be used to corroborate and refine phylogenies hypothesized by base substitutions, at least at lower taxonomic levels. At present, full use of gaps in phylogenetic reconstruction is best achieved in parsimony analysis, pending development of well-justified and generally applicable methods for incorporating indels in explicitly model-based methods.
Mol Biol Evol 2003 Jan
PMID:Evolution and phylogenetic utility of alignment gaps within intron sequences of three nuclear genes in bumble bees (Bombus). 1251 10

The effect of acclimation salinity and salinity changes on the concentration of high-energy phosphate metabolites and arginine kinase (AK) flux was examined in vivo in juvenile blue crabs using 31P-nuclear magnetic resonance (NMR). Crabs were acclimated for 7 days to a salinity of 5 or 35 per thousand and then placed in a flow apparatus that could sustain the animals while NMR spectra were acquired. Crabs were subjected to either hyperosmotic salinity changes, where an animal acclimated to 5 per thousand was exposed to a salinity of 35 per thousand, or hyposmotic changes, which involved the reciprocal exchange. Neither acclimation salinity nor salinity change had a significant effect on the concentrations of arginine phosphate, inorganic phosphate or ATP. 31P-NMR saturation transfer experiments were used to determine the effect of salinity on the forward and reverse flux of the AK reaction. There was no significant effect of acclimation salinity or salinity change on the flux rate through this reaction. This is in contrast to previous results, which showed that AK flux in isolated muscle was sensitive to prevailing osmotic conditions (Holt and Kinsey, J. Exp. Biol. 205 (2002) 1775-1785). The present study indicates that the integrated osmoregulatory capacity of the intact animal is sufficient to preserve cellular energy status and enzyme function during acute salinity changes.
Comp Biochem Physiol B Biochem Mol Biol 2003 Jul
PMID:The effects of rapid salinity change on in vivo arginine kinase flux in the juvenile blue crab, Callinectes sapidus. 1283 72

The effects of zinc on arginine kinase and its collapsed-state intermediate were studied. Both arginine kinase and the collapsed-state intermediate were inactivated in the presence of zinc, following a biphasic kinetic course. The corresponding apparent rate constants of inactivation at different zinc concentrations and conformational changes in the presence of 0.5 mM zinc were obtained. The conformational changes of arginine kinase and the collapsed-state intermediate were followed by fluorescence spectra and circular dichroism spectra. Comparison of the results for arginine kinase and the collapsed-state intermediate showed that the collapsed-state intermediate was more susceptible to zinc, which indicated that the collapsed-state intermediate was more flexible and unstable than arginine kinase. The special structure of arginine kinase might explain these diverse phenomena.
J Biochem Mol Biol 2003 Jul 31
PMID:Effects of zinc on the activity and conformational changes of arginine kinase and its intermediate. 1289 93

The clams Pseudocardium, Solen, Corbicula and Ensis possess a unique form of arginine kinase (AK) with a molecular mass of 80 kDa and an unusual two-domain structure, a result of gene duplication and subsequent fusion. These AKs also lack two functionally important amino acid residues, Asp(62) and Arg(193), which are strictly conserved in other 40-kDa AKs and are assumed to be key residues for stabilizing the substrate-bound structure. However, these AKs show higher enzyme activity. The cDNA-derived amino acid sequences of 40-kDa AKs from the blood clam Scapharca broughtonii and the oyster Crassostrea gigas were determined. While Asp(62) and Arg(193) are conserved in Scapharca AK, these two key residues are replaced by Asn and Lys, respectively, in Crassostrea AK. The native enzyme from Crassostrea and both of the recombinant enzymes show an enzyme activity similar to that of two-domain clam AKs and at least twofold higher than that of other molluskan AKs. Although the replacement of Asp(62) or Arg(193) by Gly in normal AK causes a considerable decrease in V(max) (6-15% of wild-type enzyme) and a two- to threefold increase in K(m) for arginine, the same replacement in Scapharca AK had no pronounced effect on enzyme activity. Together with the observation that bivalve AKs are phylogenetically distinct from other molluskan AKs, these results suggest that bivalve AKs have undergone a unique molecular evolution; the characteristic stabilizing function of residues 62 and 193 has been lost and, consequently, the enzyme shows higher activity than normal.
Cell Mol Life Sci 2004 Jan
PMID:Unique evolution of Bivalvia arginine kinases. 1470 58

Protein expression in unfed larvae of the cattle tick, Boophilus microplus, was characterized using gel electrophoresis and mass spectrometry in an effort to assemble a database of proteins produced at this stage of development. Soluble and insoluble proteins were extracted and resolved by two-dimensional (2D) gel electrophoresis. Twenty abundantly expressed larval proteins were selected for peptide mass mapping and for peptide sequencing by matrix-assisted laser desorption/ionization time-of-flight (MALDI-ToF) and quadrupole time-of-flight (Q-ToF) tandem mass spectrometry (MS), respectively. Only one protein, tropomyosin, was unequivocally identified from its peptide mass map. Ten proteins were assigned putative identities based on BLAST searching of heterologous databases with peptide sequences. These included a cytoskeletal protein (troponin I), multiple cuticular proteins, a glycine-rich salivary gland-associated protein and proteins with a presumed housekeeping role (arginine kinase, a high-mobility group protein and a small heat shock protein). Eight additional proteins were identified by searching translated open reading frames of a B. microplus EST database (unpublished): putative fatty-acid binding protein, thioredoxin, glycine-rich salivary gland protein and additional cuticular proteins. One remaining protein was not identifiable, suggesting it may be a novel molecule. The ongoing assembly of this database contributes to our understanding of proteins expressed by the tick and provides a resource that can be mined for molecules that play a role in tick-host interactions.
Insect Biochem Mol Biol 2005 Feb
PMID:Proteome analysis of abundantly expressed proteins from unfed larvae of the cattle tick, Boophilus microplus. 1568 Dec 24

Previous phylogenetic work on the Hawaiian bees of the genus Hylaeus, based on mitochondrial DNA and morphology, appeared to support a recent origin for the group, but support for the resulting tree was weak. Four nuclear genes with varying evolutionary rates -- arginine kinase, EF-1alpha, opsin, and wingless -- were sequenced for a reduced taxon set in an attempt to find one or more data set that would provide better support. All showed very low variation (<2%) in the ingroup. Comparison among genes revealed a much higher than expected rate of evolution in mtDNA, especially at first and second positions. While the data from the nuclear genes showed insufficient variation for phylogenetic analysis, the strong sequence similarity among the Hawaiian species supports the previous hypothesis of a recent origin for the group.
Mol Phylogenet Evol 2007 Jun
PMID:Low nuclear DNA variation supports a recent origin of Hawaiian Hylaeus bees (Hymenoptera: Colletidae). 1704 77

In response to hypoxia at PO(2) 1.3-1.7 mg/L for 6 h, the kuruma prawn Marsupenaeus (Penaeus) japonicus showed a dramatic decrease in phosphoarginine storage in muscle, with normal levels restored during 4-h post-hypoxic recovery. Large stores of muscle glycogen only decreased between 4 and 6 h during hypoxia, but greatly diminished during recovery. Muscle ATP levels and energy charge decreased only slightly under hypoxia. Lactate levels increased slightly during hypoxia and promptly returned to control levels during recovery. These data indicate that phosphoarginine works in muscle as an ATP buffer during hypoxia and glycogen is utilized as an energy source during recovery. Under hypoxia, up- and down-regulated proteins were identified after 2D electrophoresis and partial sequences were obtained after protease digestion. Fructose bisphosphate aldolase was down-regulated during hypoxia, suggesting the suppression of glycolysis under hypoxia. Several partial sequences from three protein spots up-regulated under hypoxia were all assigned to arginine kinase, suggesting the existence of several isoforms of arginine kinase in the muscle of M. japonicus. This arginine kinase up-regulation under hypoxia may indicate a provision for oxygen re-supply after anaerobiosis. This is consistent with the prompt replenishment of phosphoarginine stores during recovery from hypoxia.
Comp Biochem Physiol A Mol Integr Physiol 2007 Jan
PMID:Metabolic responses and arginine kinase expression under hypoxic stress of the kuruma prawn Marsupenaeus japonicus. 1708 99


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