Protein hydrolysate from miscellaneous fish

A method to prepare fish protein hydrolysate from miscellaneous fish obtained as by catch from shrimp trawlers is outlined. Effect of temperature and concentration of enzyme papain on the yield of hydrolysates has been determined. It is seen that within 30 min at 55°C and pH 6.5 fish proteins can be effectively solubilized, provided the nitrogen content of the enzyme (activity 10 units/mg enzyme) and substrate are maintained in the ratio 1:30. This hydrolysate possesses the best amino acid pattern compared to those obtained after hydrolysis for 60 to 180 min.

ABI1 and PP2CA Phosphatases Are Negative Regulators of Snf1-Related Protein Kinase1 Signaling in Arabidopsis

Plant survival under environmental stress requires the integration of multiple signaling pathways into a coordinated response, but the molecular mechanisms underlying this integration are poorly understood. Stress-derived energy deprivation activates the Snf1-related protein kinases1 (SnRK1s), triggering a vast transcriptional and metabolic reprogramming that restores homeostasis and promotes tolerance to adverse conditions. Here, we show that two clade A type 2C protein phosphatases (PP2Cs), established repressors of the abscisic acid (ABA) hormonal pathway, interact with the SnRK1 catalytic subunit causing its dephosphorylation and inactivation. Accordingly, SnRK1 repression is abrogated in double and quadruple pp2c knockout mutants, provoking, similarly to SnRK1 overexpression, sugar hypersensitivity during early seedling development. Reporter gene assays and SnRK1 target gene expression analyses further demonstrate that PP2C inhibition by ABA results in SnRK1 activation, promoting SnRK1 signaling during stress and once the energy deficit subsides. Consistent with this, SnRK1 and ABA induce largely overlapping transcriptional responses. Hence, the PP2C hub allows the coordinated activation of ABA and energy signaling, strengthening the stress response through the cooperation of two key and complementary pathways.

Riboflavin carrier protein from carp (C. carpio) eggs: comparison with avian riboflavin carrier protein

A protein exhibiting immunological cross-reactivity with the chicken egg-white riboflavin carrier protein was detected by radioimmunoassay in the eggs and serum of the fresh water fish Cyprinus carpio and subsequently purified to homogeneity by use of affinity chromatography. Fish riboflavin carrier protein resembled chicken riboflavin carrier protein with respect to most of its physicochemical characteristics. The major epitopes of chicken riboflavin carrier protein were shown to be conserved in the fish protein as probed with monoclonal antibodies to the avian vitamin carrier.

An appraisal of the efficiency of fish marketing system in Lake Chad basin

Fish products from the Chad Basin Lake play important role in meeting fish protein needs of Nigeria: they contribute not less than 25% of the total domestic fish supply and are significant in determining the availability of processed products and reduction of post-harvest losses. Processors, marketers and consumers are the major actors in appraising a marketing system. The results show that most sellers (4-7.5%) are within the age range of 30-39 years. Desires for more earnings led the markets to diversify their business activities to food stuff trading (37.5%), dried meat/livestock sales (37.5%), farming (12.5%), and transportation (12.5%). 65% of traders dispose off their products mostly in the mornings and evenings, 70% of the products are sold smoked while 50% of products are sold to individual consumers. Lake Chad fish products have a long distribution chain. There is also a high degree of buyers and sellers concentration in the primary fish markets and secondary (urban) markets. The products have a vertical regional movement with southern traders (82.5%) dominating the business, thus making the products popular all over Nigeria. Product differentiation with imperfect pricing policy is common occurrence. Lake Chad fish marketing system has distortions that impede its efficiency, recommendations are made on how to ensure a better efficiency of the system

Approaches towards increasing domestic fish production in Nigeria: "Fish for All"

The paper critically examines the factors militating against domestic fish production in artisanal, aquacultural and industrial sectors in Nigeria. Approaches towards increasing fish production from all these sectors were suggested and discussed in line with the National Economic Empowerment Development Strategy (NEEDS) and Poverty Alleviation programmes of the Federal Government to meet the nation's fish protein requirement within the next three years

Protein from blanch liquor

A process is described for isolation of edible protein from blanch liquor, which is discarded as a waste at present from prawn canning factories. The protein isolated is colourless and odourless and contains an appreciable amount of salt from the brine used for blanching prawns. It is comparable to fish protein concentrate in amino acid composition.

Protein concentrate from shark

A simple method of leaching the minced muscle with water repeatedly followed by cooking, pressing, drying the cake and powdering has been described for the preparation of fish protein concentrate (FPC) from shark without the use of solvents. The FPC thus prepared had high protein content and was completely free of urea. It contained all the essential amino acids in a balanced proportion with high lysine content and had a storage life up to 12 months. This product can be used for the fortification of bread, biscuits and chappathis respectively at 10, 5 and 2% levels.

Increased insulin-stimulated Akt pSer473 and cytosolic SHP2 protein abundance in human skeletal muscle following acute exercise and short-term training.

The purpose of the present study was to determine in human skeletal muscle whether a single exercise bout and 7 days of consecutive endurance (cycling) training 1) increased insulin-stimulated Akt pSer473and 2) altered the abundance of the protein tyrosine phosphatases (PTPases), PTP1B and SHP2. In healthy, untrained men (n = 8; 24 ± 1 yr), glucose infusion rate during a hyperinsulinemic euglycemic clamp, when compared with untrained values, was not improved 24 h following a single 60-min bout of endurance cycling but was significantly increased (~30%; P < 0.05) 24 h following completion of 7 days of exercise training. Insulin-stimulated Akt pSer473was ~50% higher (P < 0.05) 24 h following the acute bout of exercise, with this effect remaining after 7 days of training (P < 0.05). Insulin-stimulated insulin receptor and insulin receptor substrate-1 tyrosine phosphorylation were not altered 24 h after acute exercise and short-term training. Insulin did not acutely regulate the localization of the PTPases, PTP1B or SHP2, although cytosolic protein abundance of SHP2 was increased (P < 0.05; main effect) 24 h following acute exercise and short-term training. In conclusion, insulin-sensitive Akt pSer473and cytosolic SHP2 protein abundance are higher after acute exercise and short-term training, and this effect appears largely due to the residual effects of the last bout of prior exercise. The significance of exercise-induced alterations in cytosolic SHP2 and insulin-stimulated Akt pSer473on the improvement in insulin sensitivity requires further elucidation.

Anti-hypertensive dietary supplement derived from Salmo or Oncorhynchus protein hydrolysates

An anti-hypertensive fish protein hydrolysate is provided, wherein the fish is of the genus Salmo or Oncorhynchus, and wherein the fish protein hydrolysate that is prepared using bacillolysm from Bacillus stearothermophilus comprises at least one peptide selected from the group consisting of Leu-Ala-Phe, Leu-Thr-Phe, Ile-Ile-Phe, Leu-Ala-Tyr, Ile-Ala-Tyr, Val-Phe-Tyr, Tyr-Ala-Tyr, Val-Leu-Trp, Ile-Ala-Trp, Tyr- Ala-Leu and Tyr-Asn-Arg Method of making and methods for using such fish protein hydrolysates are also provided.

Anti-hypertensive dietary supplement derived from salmo or oncorhynchus protein hydrolysates

An anti-hypertensive fish protein hydrolysate is provided, wherein the fish is of the genus Salmo or Oncorhynchus, and wherein the fish protein hydrolysate that is prepared using bacillolysm from Bacillus stearothermophilus comprises at least one peptide selected from the group consisting of Leu-Ala-Phe, Leu-Thr-Phe, Ile-Ile-Phe, Leu-Ala-Tyr, Ile-Ala-Tyr, Val-Phe-Tyr, Tyr-Ala-Tyr, Val-Leu-Trp, Ile-Ala-Trp, Tyr- Ala-Leu and Tyr-Asn-Arg Method of making and methods for using such fish protein hydrolysates are also provided.

The protein kinases of Caenorhabditis elegans: A model for signal transduction in multicellular organisms

Caenorhabditis elegans should soon be the first multicellular organism whose complete genomic sequence has been determined. This achievement provides a unique opportunity for a comprehensive assessment of the signal transduction molecules required for the existence of a multicellular animal. Although the worm C. elegans may not much resemble humans, the molecules that regulate signal transduction in these two organisms prove to be quite similar. We focus here on the content and diversity of protein kinases present in worms, together with an assessment of other classes of proteins that regulate protein phosphorylation. By systematic analysis of the 19,099 predicted C. elegans proteins, and thorough analysis of the finished and unfinished genomic sequences, we have identified 411 full length protein kinases and 21 partial kinase fragments. We also describe 82 additional proteins that are predicted to be structurally similar to conventional protein kinases even though they share minimal primary sequence identity. Finally, the richness of phosphorylation-dependent signaling pathways in worms is further supported with the identification of 185 protein phosphatases and 128 phosphoprotein-binding domains (SH2, PTB, STYX, SBF, 14-3-3, FHA, and WW) in the worm genome.

Vimentin Dephosphorylation by Protein Phosphatase 2A Is Modulated by the Targeting Subunit B55

The intermediate filament protein vimentin is a major phosphoprotein in mammalian fibroblasts, and reversible phosphorylation plays a key role in its dynamic rearrangement. Selective inhibition of type 2A but not type 1 protein phosphatases led to hyperphosphorylation and concomitant disassembly of vimentin, characterized by a collapse into bundles around the nucleus. We have analyzed the potential role of one of the major protein phosphatase 2A (PP2A) regulatory subunits, B55, in vimentin dephosphorylation. In mammalian fibroblasts, B55 protein was distributed ubiquitously throughout the cytoplasm with a fraction associated to vimentin. Specific depletion of B55 in living cells by antisense B55 RNA was accompanied by disassembly and increased phosphorylation of vimentin, as when type 2A phosphatases were inhibited using okadaic acid. The presence of B55 was a prerequisite for PP2A to efficiently dephosphorylate vimentin in vitro or to induce filament reassembly in situ. Both biochemical fractionation and immunofluorescence analysis of detergent-extracted cells revealed that fractions of PP2Ac, PR65, and B55 were tightly associated with vimentin. Furthermore, vimentin-associated PP2A catalytic subunit was displaced in B55-depleted cells. Taken together these data show that, in mammalian fibroblasts, the intermediate filament protein vimentin is dephosphorylated by PP2A, an event targeted by B55.

Vacuole Fusion Regulated by Protein Phosphatase 2C in Fission Yeast

The gene ptc4+ encodes one of four type 2C protein phosphatases (PP2C) in the fission yeast Schizosaccharomyces pombe. Deletion of ptc4+ is not lethal; however, Δptc4 cells grow slowly in defined minimal medium and undergo premature growth arrest in response to nitrogen starvation. Interestingly, Δptc4 cells are unable to fuse vacuoles in response to hypotonic stress or nutrient starvation. Conversely, Ptc4 overexpression appears to induce vacuole fusion. These findings reveal a hitherto unrecognized function of type 2C protein phosphatases: regulation of vacuole fusion. Ptc4 localizes in vacuole membranes, which suggests that Ptc4 regulates vacuole fusion by dephosphorylation of one or more proteins in the vacuole membrane. Vacuole function is required for the process of autophagy that is induced by nutrient starvation; thus, the vacuole defect of Δptc4 cells might explain why these cells undergo premature growth arrest in response to nitrogen starvation.

Phosphorylation and microtubule association of the Opitz syndrome protein mid-1 is regulated by protein phosphatase 2A via binding to the regulatory subunit α4

Opitz syndrome (OS) is a human genetic disease characterized by deformities such as cleft palate that are attributable to defects in embryonic development at the midline. Gene mapping has identified OS mutations within a protein called Mid1. Wild-type Mid1 predominantly colocalizes with microtubules, in contrast to mutant versions of Mid1 that appear clustered in the cytosol. Using yeast two-hybrid screening, we found that the α4-subunit of protein phosphatases 2A/4/6 binds Mid1. Epitope-tagged α4 coimmunoprecipitated endogenous or coexpressed Mid1 from COS7 cells, and this required only the conserved C-terminal region of α4. Localization of Mid1 and α4 was influenced by one another in transiently transfected cells. Mid1 could recruit α4 onto microtubules, and high levels of α4 could displace Mid1 into the cytosol. Metabolic 32P labeling of cells showed that Mid1 is a phosphoprotein, and coexpression of full-length α4 decreased Mid1 phosphorylation, indicative of a functional interaction. Association of green fluorescent protein–Mid1 with microtubules in living cells was perturbed by inhibitors of MAP kinase activation. The conclusion is that Mid1 association with microtubules, which seems important for normal midline development, is regulated by dynamic phosphorylation involving MAP kinase and protein phosphatase that is targeted specifically to Mid1 by α4. Human birth defects may result from environmental or genetic disruption of this regulatory cycle.

Study of the role of retinoblastoma protein in terminal differentiation of murine erythroleukemia cells.

Hexamethylenebisacetamide-induced terminal differentiation of Friend virus-transformed murine erythroleukemia (MEL) cells can be inhibited by okadaic acid, an inhibitor of type 1 and type 2A protein phosphatases. The inhibition is shown to be correlated with prevention of dephosphorylation of retinoblastoma protein (pRB) in cells and bypass of G1 prolongation in the cell cycle. These results suggest that pRB-mediated G1 prolongation is necessary for MEL cells to commit to terminal differentiation. However, further experiments demonstrate that the simple cell cycle exit is not sufficient for commitment to terminal differentiation. Induction of dephosphorylation of pRB and subsequent G1 prolongation by forskolin does not lead MEL cells to differentiate. Additional pRB has been expressed in MEL cells by transfection with a neo-resistant plasmid containing RB cDNA under the control of a cytomegalovirus promoter. Exogenously expressed pRB is hyperphosphorylated in logarithmically growing MEL cells without any noticeable change in growth rate between the transfected cell line and the parental cell line. This result suggests that pRB in MEL cells is regulated by protein kinases and protein phosphatases and not by transcription.