Ion-selective enrichment of tyrosine-sulfated peptides from complex protein digests.

We have developed a novel procedure for concentrating sulfated peptides, as a front end to mass spectrometric analysis, based on ion-selective interaction of sulfate ions with anion exchangers. Ions with a higher charge and smaller solvated ion radius, such as sulfate ions, have higher retention in an ion exchanger due to their greater degree of coulombic interactions. We tested the effectiveness of this approach for enrichment and identification of sulfated peptides using a tryptic digest of bovine serum albumin spiked with model sulfated peptide (molar ratio 20:1) and using a tryptic digest of bovine fibrinogen.

Effects of diet on the incidence of dental pathology in free living caviomorph rodents.

Examinations of dental pathology in five genera of caviomorph rodents found caries and periodontal disease in both grazers and frugivores, although the prevalence differed between the two groups. Caries was more prevalent in the frugivores (10.5-19.

Mechanism of reduction in the number of the circulating hemocytes in the Pseudaletia separata host parasitized by Cotesia kariyai.

Larval endoparasitoids can avoid the immune response of the host by the function of polydnavirus (PDV) and venom. PDV infects hemocytes and affects the hemocyte function of the host. In this paper, we investigated how PDV and venom affect the hemocyte population of the host.

The function of a trypsin-like enzyme in the saliva of Euplectrus separatae larvae.

Larvae of the gregarious ectoparasitoid, Euplectrus separatae, a species that parasitizes Pseudaletia separata, migrate from the dorsal to the ventral side of the host larva for pupation 7 days after parasitization. The parasitized host larvae die after the migration. The body mass of the parasitoid larvae increases while that of the host larva drastically decreases.

Dental and skull anomalies in feral coypu, Myocastor coypus.

The study of dental and skull abnormalities of wild animals can provide useful information for comparative studies. Although studies of captive and laboratory rodents have been published, there are few reports on wild populations. We examined anomalous and pathological conditions in the skulls of feral coypus, Myocastor coypus, from Aichi Prefecture, Japan, and identified the types of abnormality.

Chemical nursing: phytosulfokine improves genetic transformation efficiency by promoting the proliferation of surviving cells on selective media.

The relative growth rate of plant cells in vitro is considerably affected by initial cell density. This troublesome effect has interfered with the establishment of efficient plant cell culture systems, especially when only a small number of cells are expected to survive, such as in the genetic transformation of cells under antibiotic selection. To improve the recovery of antibiotic-resistant cells, we examined the use of the peptide plant hormone phytosulfokine (PSK), which has been shown to promote cellular growth and development in vitro.

Venom of Euplectrus separatae causes hyperlipidemia by lysis of host fat body cells.

Although the lepidopteran larva Pseudaletia separata is attacked by the gregarious ectoparasitoid Euplectrus separatae, it continues to feed and grow. Lipid concentration in the hemolymph of the parasitized host was higher than that of the nonparasitized host from 3 to 8 days after parasitization. Artificial injection of parasitoid venom also elevated lipid concentration in the host hemolymph.

Correlation between concentration of hemolymph nutrients and amount of fat body consumed in lightly and heavily parasitized hosts (Pseudaletia separata).

Two states of parasitization in the Pseudaletia separata-Cotesia kariyai system were examined: one that was lightly parasitized and one that was heavily parasitized. We predicted that the consumption of fat body and hemolymph nutrients depends on the number of parasitoid larvae in the host. Lightly parasitized hosts (average clutch size+/-S.

Ligand-receptor pairs in plant peptide signaling.

Extensive studies on plant signaling molecules over the past decade indicate that plant cell-to-cell communication, as is the case with animal systems, makes use of small peptide signals and specific receptors. To date, four peptide-ligand-receptor pairs have been identified and shown to be involved in a variety of processes. Systemin and phytosulfokine (PSK), the first and second signaling peptides identified in plants, were isolated by biochemical purification based on their biological activities.

Similar polydnavirus genes of two parasitoids, Cotesia kariyai and Cotesia ruficrus, of the host Pseudaletia separata.

Endoparasitoids can achieve successful development in suitable hosts with the help of polydnavirus (PDV), if it is injected into the host with their eggs at parasitization. It was predicted that different species of endoparasitoids that parasitize the same host successfully use a similar system to avoid the defense reaction from the host. In this study, we identified similar PDVs genes.

Larvae of an endoparasitoid, Cotesia kariyai (Hymenoptera: Braconidae), feed on the host fat body directly in the second stadium with the help of teratocytes.

Larvae of a gregarious endoparasitoid, Cotesia kariyai (Watanabe), grew rapidly during the second stadium in the host. The fat body of a Pseudaletia host parasitized by C. kariyai was completely consumed by 10 d, just before larval emergence.

Venom of ectoparasitoid, Euplectrus sp near plathypenae (Hymenoptera: Eulophidae) regulates the physiological state of Pseudaletia separata (Lepidoptera: Noctuidae) host as a food resource.

Euplectrus sp. near plathypenae is an ectoparasitoid that can parasitize from 3rd to day 0-6th instar Pseudaletia separata. The developmental period of the parasitoid from the egg to the pupal stage is about 13 days.

An LRR receptor kinase involved in perception of a peptide plant hormone, phytosulfokine.

The sulfated peptide phytosulfokine (PSK) is an intercellular signal that plays a key role in cellular dedifferentiation and proliferation in plants. Using ligand-based affinity chromatography, we purified a 120-kilodalton membrane protein, specifically interacting with PSK, from carrot microsomal fractions. The corresponding complementary DNA encodes a 1021-amino acid receptor kinase that contains extracellular leucine-rich repeats, a single transmembrane domain, and a cytoplasmic kinase domain.

Peptide signals and their receptors in higher plants.

At least four peptides play a vital role in plant cell-cell communication by means of their specific receptors. Two of these receptors have been identified as receptor kinases, which form a large family of receptor molecules in plants. These findings highlight the significance of receptor-mediated peptide signaling in various physiological events in plants, and predict the existence of further peptide-signal-interacting receptor kinases.

Diversity of Arabidopsis genes encoding precursors for phytosulfokine, a peptide growth factor.

Phytosulfokine-alpha (PSK-alpha), a unique plant peptide growth factor, was originally isolated from conditioned medium of asparagus (Asparagus officinalis) mesophyll cell cultures. PSK-alpha has several biological activities including promoting plant cell proliferation. Four genes that encode precursors of PSK-alpha have been identified from Arabidopsis.

Pattern of X-Y chromosome pairing in the Taiwan vole, Microtus kikuchii.

Pairing of X and Y chromosomes at meiotic prophase and the G- and C-banding patterns and nucleolar organizer region (NOR) distribution were analyzed in Microtus kikuchii. M. kikuchii is closely related to M.

Molecular cloning and characterization of OsPSK, a gene encoding a precursor for phytosulfokine-alpha, required for rice cell proliferation.

We previously characterized an OsPSK cDNA encoding a precursor of phytosulfokine-alpha (PSK-alpha), a peptide plant growth factor. Southern blot analysis suggested that OsPSK is a single-copy gene in rice, which we have isolated and characterized. The OsPSK gene consists of one large intron and two exons.

Cotesia kariyai teratocytes: growth and development.

The growth and development of teratocytes was examined in the Cotesia kariyai-Pseudaletia separata system. Cotesia kariyai embryos released an average of 163 teratocytes at the time of hatching, 3.5 days after oviposition.

Phytosulfokine-alpha, a peptide growth factor found in higher plants: its structure, functions, precursor and receptors.

Phytosulfokine-alpha, a sulfated pentapeptide growth factor universally found in both monocotyledons and dicotyledons, strongly promotes proliferation of plant cells in culture. It is similar to animal polypeptide hormones in that it is processed from a larger precursor, preprophytosulfokine, although the putative processing sites do not conform to consensus sequences for endoproteolytic processing sites flanking animal prohormones. Like the animal preprohormones, preprophytosulfokine also has a signal peptide at the N-terminus for targeting to secretory pathways.

In vivo gene electroporation confers strong transient expression of foreign genes in the chicken testis.

To attain foreign gene expression in vivo in the testis of living chickens, chloramphenicol acetyltransferase (CAT) and firefly luciferase reporter genes were transfected by electroporation (EP). Bioluminescence imaging indicated clear expression of the luciferase reporter gene localized in and around the injection site of the chicken testis. The CAT activity decreased sharply from 7 to 14 d posttransfection (P < 0.

A rapid and efficient system of Agrobacterium infection-mediated transient gene expression in rice Oc cells and its application for analysis of the expression and antisense suppression of preprophytosulfokine, a precursor of phytosulfokine-a, encoded by OsPSK gene.

A rapid and efficient system for Agrobacterium infection-mediated transient gene expression in rice has been developed. Using this system, transient expression of preprophytosulfokine, a precursor of phytosulfokine-a, encoded by OsPSK gene was analyzed. The results suggest that the Agrobacterium infection-mediated transient gene expression system is as efficient in rice Oc cells as in tobacco BY-2 cells and might be useful for rapid analysis not only of foreign gene expression, but also of antisense gene suppression.

120- and 160-kDa receptors for endogenous mitogenic peptide, phytosulfokine-alpha, in rice plasma membranes.

Plant cells in culture secrete a sulfated peptide named phytosulfokine-alpha (PSK-alpha), and this peptide induces the cell division and/or cell differentiation by means of specific high and low affinity receptors. Putative receptor proteins for this autocrine type growth factor were identified by photoaffinity labeling of plasma membrane fractions derived from rice suspension cells. Incubation of membranes with a photoactivable (125)I-labeled PSK-alpha analog, [N(epsilon)-(4-azidosalicyl)Lys(5)]PSK-alpha (AS-PSK-alpha), followed by UV irradiation resulted in specific labeling of 120- and 160-kDa bands in SDS-polyacrylamide gel electrophoresis.

Oryza sativa PSK gene encodes a precursor of phytosulfokine-alpha, a sulfated peptide growth factor found in plants.

Phytosulfokine-alpha [PSK-alpha, Tyr(SO(3)H)-Ile-Tyr(SO(3)H)-Thr-Gln], a sulfated mitogenic peptide found in plants, strongly promotes proliferation of plant cells in culture at very low concentrations. Oryza sativa PSK (OsPSK) cDNA encoding a PSK-alpha precursor has been isolated. The cDNA is 725 base pairs long, and the 89-aa product, preprophytosulfokine, has a 22-aa hydrophobic region that resembles a cleavable leader peptide at its NH(2) terminus.

Protein transport within the plant cell endomembrane system: an update.

The secretory pathway plays a central role in plant development and morphogenesis. Storage protein deposition, plant cell division and the expansion of the plasma membrane and extracellular matrix all require the synthesis and trafficking of membranes, proteins and polysaccharides through this network of organelles. Increasing evidence demonstrates that the plant secretory pathway is more complex than previously appreciated and that its formation and maintenance are guided/regulated by many different mechanisms.

Preparation and Characterization of Fully Active Biotinylated Analogs of Phytosulfokine-α.

We report the preparation of biotinylated analogs of phytosulfokine-α (Tyr(SO3H)-Ile-Tyr(SO3H)-Thr-Gln; PSK-α), an endogenous peptide growth factor in plants. Because the modification of the N-terminal amino group leads to significant loss of the activities, a Lys residue was incorporated in the C-terminal region of PSK-α, and its e amino group was reacted with biotinylation reagent. Results of the binding assay showed that [N(ε)-(biotinyl)Lys(5)]PSK-α retained the same binding activity and mitogenic activity as that of native PSK-α.