Comparative examination of the soluble muscle proteins of two species of angler-fish (Lophiidae): Lophius piscatorius (L.) and Lophius budegassa (Spinola)

• 1.1. A comparative examination of sarcoplasmic proteins of the two nominal European species of angler-fish, Lophius piscatorius and L. budegassa was carried out using isoelectric focusing techniques.
• 2.2. Two protein bands differing in isoelectric point proved diagnostic for L. budegassa (pI 4.40 and pI 5.75) while a third characterized L. piscatorius (pI 4.65).
• 3.3. These species-specific protein profiles provide a method of species discrimination independent of morphological criteria.
• 4.4. Within-species heterogeneity of banding pattern suggested the presence of polymorphic gene loci of potential use in studies of population structure.

     

Phosphorylation of Escherichia coli proteins during the SOS response

• 1.1. The phosphorylation of Escherichia coli proteins was analyzed comparatively before and after induction of the SOS response in a temperature-sensitive mutant strain.
• 2.2. The presence of phosphorylated proteins was evidenced by gel electrophoresis and autoradiography after labelling with radioactive orthophosphate in vivo or radioactive adenosine triphosphate in vitro.
• 3.3. Significant changes in the intensity of protein labelling were observed upon induction of the SOS functions: six proteins were found to be more phosphorylated while two others were less phosphorylated. Moreover, five additional proteins appeared to become phosphorylated exclusively during the SOS response. The molecular mass and isoelectric point of these various proteins were determined.
• 4.4. For most proteins, the changes in the pattern of protein phosphorylation were concomitant with variations in the amount of protein synthesized.
• 5.5. The changes in the pattern of phosphoproteins observed during the SOS response were not due to the temperature shift required experimentally for expressing the SOS phenotype.
• 6.6. Phosphorylation was found to be catalyzed by protein kinases that modify amino acid residues at hydroxyl groups in protein substrates.
• 7.7. Both in vivo and in vitro studies brought evidence that neither RecA nor LexA, the two key regulatory proteins of the SOS functions, were capable of undergoing phosphorylation.

     

Isolation and characterization of the plasma membrane from slime-forming,encapsulated Streptococcus cremoris

• 1.1. The plasma membrane of slime-forming, encapsulated Streptococcus cremoris from “viili” was isolated in hypotonie conditions in the presence of lysozyme (EC 3.2.1.17) using density gradient centrifugation as the last purification step.
• 2.2. The membrane yield was 15.8% of wet weight cells and the preparation contained 64.4% protein. 19.1% carbohydrate, 5.8% aminosugars, 5.1% RNA and 0.07% DNA.
• 3.3. Buffered 1% (w/v) Triton X-100 solubilized 33.6% of membrane proteins. The number of polypeptides detected by SDS-polyacrylamide gel electrophoresis was 59 when the membrane was isolated without a protease inhibitor and 44 in the presence of a protease inhibitor.
• 4.4. The molecular weights of the polypeptides varied from 13,500 to 100,000.
• 5.5. Ultrathin-layer electrofocusing analysis revealed the range of protein pi values to be between 3.50 and 5.85 concerning 77.3% of proteins and between pI 5.85 and 8.15 concerning 18.2% of proteins.
• 6.6. The isoelectric point of the only basic protein component was 9.3.

     

Bioassay and proteolytic activity of digestive enzymes from octopus saliva

• 1.1. A bioassay for octopus saliva, based on detachment of crab dactylopodite flexor muscle under standard conditions, has been developed.
• 2.2. There is a direct relationship between increasing caseinolytic activity of saliva from Eledone cirrhosa and decreasing muscle detachment time.
• 3.3. Fractionation of saliva, using preparative isoelectric focusing, shows that muscle releasing activity is restricted to fractions containing proteins with high isoelectric points and maximum caseinase activity.
• 4.4. It is concluded that proteolytic enzyme(s) in octopus saliva selectively release crab muscle from attachment to the carapace.

     

Study of the soluble lens proteins from fifteen sparidae species

• 1.1. Soluble eye lens proteins of fifteen different Sparidae species were analysed.
• 2.2. Species-specific electrophoretic and isoelectric focusing patterns were found.
• 3.3. Significant differences in the distribution of β and γ-crystallin protein components were noted for all species.
• 4.4. These data suggest that the Sparidae family may be a heterogenenous taxonomic group encompassing considerable genetic diferences and with different evolutionary histories.

     

Effects of niacin deficiency on the relative turnover rates of proteins in various tissues of Japanese quail

• 1.1. The effects of niacin deficiency on the relative turnover rates of proteins in various tissues of Japanese quail were investigated.
• 2.2. The level of liver NAD was not affected by niacin deficiency whereas the level of pectoral muscle NAD was markedly reduced.
• 3.3. In all dietary treatments the liver had the highest turnover rates of proteins, heart and brain had intermediate rates, and pectoral muscle had the lowest rates.
• 4.4. Relative turnover rates of proteins in all tissues (particularly pectoral muscle) of the niacin deficient group were significantly higher than those of pair-fed control group, although there were no significant differences in turnover rate between pair-fed control and control groups.
• 5.5. The high turnover rate of proteins in niacin deficiency was primarily attributed to enhanced degradation rate of proteins rather than enhanced synthesis rate of proteins.
• 6.6. Optical density scanning (or densitometric) of water-soluble pectoral muscle proteins separated by isoelectric focusing revealed several additional minor protein bands between major protein bands in the niacin deficient group which were more pronounced in the acidic region of the gel.
• 7.7. These results suggest that proteins with a low pI value in pectoral muscle of the niacin deficient animal are highly sensitive to protein degradation.

     

The effects of chronic metabolic acidosis on patterns of protein synthesis in rat renal cortex

• 1.1. In the rat chronic metabolic acidosis increases the net synthesis of 17 renal cortex proteins by amounts ranging from 1.5 to 4.5-fold.
• 2.2. These proteins have molecular weights between 13,000 and 42,000 and isoelectric points between approximately 5.5 and 7.0.
• 3.3. No new proteins not also present in normal animals are detected in renal cortex samples from acidotic animals.
• 4.4. Three proteins undergo substantial reductions in their net synthetic rates in chronic metabolic acidosis.
• 5.5. On the basis of their physical properties and similar alterations in net synthetic rate in acidosis some of these proteins appear to be closely related and may be coordinately expressed in the rat kidney.

     

Locust vitellogenin receptor: an acidic glycoprotein with N- and O-linked oligosaccharides

• 1.1. The locust vitellogenin (VTG) receptor which is embedded in oocyte plasma membranes is a glycoprotein.
• 2.2. With various lectins oligosaccharide units have been identified, among them neuraminic acid linked to Gal or GalNAc, mannose chains, Gal linked to GalNAc or GlcNAc and fucose linked to GlcNAc.
• 3.3. With specific enzymes it could be shown that mannose and most other oligosaccharides are O-linked while others like fucose are N-linked.
• 4.4. Enzymatic removal of all O-linked carbohydrates resulted in a drop of the molecular mass of the receptor protein from 200,000 to 110,000.
• 5.5. A total of N- and O-linked oligosaccharides of 54% was calculated.
• 6.6. The isoelectric point of the receptor was found to be at pH 3.4 increasing slightly after removal of neuraminic acid.
• 7.7. Removal of neuraminic acids destroyed the binding ability for VTG.

     

Possible involvement of adenylylation in the modification of a 26 kDa protein in rat parotid acinar cells

• 1.1. Adenylylation, a posttranslational modification of proteins, was investigated in saponin-permeabilized acinar cells of the rat parotid gland.
• 2.2. When cells were incubated with [2,8-³H]ATP, several proteins, including a 26 kDa protein in the particulate fraction, were labeled.
• 3.3. Upon incubation of cells with [α-³²P]ATP in the presence of cAMP and 3-isobutyl-lmethylxanthine, ³²P-labeling of the 26 kDa protein was observed.
• 4.4. After treatment with snake venom phosphodiesterase, [³²P]AMP was released from the 26kDa protein. Such release was not observed when cells were labeled with [γ-³²P]ATP.
• 5.5. The ³²P-labeling pattern of proteins with [α-³²P]ATP was clearly different from that with [adenylate-³²P]NAD⁺.
• 6.6. The results suggest that the 26 kDa protein is one of the adenylylation substrates in rat parotid acinar cells.

     

Protein characterization of sexually mature and immature forms of Schistosoma mansoni

• 1.1. Protein composition of different stages of Schistosoma mansoni was compared using specific antisera, 2D polyacrylamide gel electrophoresis and 14-C-leucine incorporation into proteins.
• 2.2. Major qualitative differences were detected when an anti-membrane antiserum was used.
• 3.3. 2D gel electrophoresis showed that the protein composition varied when mature and immature females were compared, whereas no differences were noted when mature and immature male worms were compared.
• 4.4. Experiments measuring protein synthesis by the different schistosome stages confirmed that upon maturation, only the female schistosomes displayed qualitative differences.
• 5.5. The protein pattern of the male schistosomes did not vary significantly as a function of development.

     

Soluble eye lens proteins of Italian ictalurids

• 1.1. Soluble eye lens proteins of three species of Italian freshwater ictalurids were analyzed: Ictalurus sp., I. nebulosus marmoratus and I. punctatus.
• 2.2. The electrophoretic and isoelectric focusing patterns were compared.
• 3.3. Both techniques revealed species-specific patterns. I. sp. and I. nebulosus marmoratus exhibited very similar patterns, I. punctatus a quite distinct one.
• 4.4. Some hypotheses warranting further investigation of the subject were proposed.

     

Binding of steroidal sex hormones by supernatant from Trichostrongylus colubriformis (Nematoda)

• 1.1. Binding of sex steroids by a supernatant from the ruminant nematode Trichostrongylus colubriformis was determined by isoelectric focusing of radiolabeled hormones.
• 2.2. Progesterone was recovered from the gels at pH 6.3 and 7.7 while 17-β-estradiol was found at pH 5.9 and 6.5. Testosterone was focused at pH 4.3. The binding of hormones was different in the sexes of nematode.
• 3.3. T. colubriformis may contain binding proteins and/or receptors for sex steroids.

     

Detection of 180 kDa proteins in electroplax sodium channel preparations

• 1.1. Co-isolating proteins (M_r 170,000–220,000) from sodium channel preparations made from the electric organ of the electric eel (Electrophorus electricus) were detected on Western blots using monoclonal a antibodies.
• 2.2. Similar protein patterns were seen on immunoblots containing immunoprecipitated protein from eel muscle and brain tissues but not heart.
• 3.3. These co-isolating proteins could be separated from the mature TTX-sensitive channel protein (M_r 280,000) using a lentil lectin-Sepharose column.
• 4.4. The 180 kDa proteins do not appear to be channel-related and can be detected as contaminants in electroplax sodium channel preparations using the monoclonal antibodies described here.

     

Electrophoretic characterization of a hybrid between Eretmochelys imbricata and Caretta caretta (Cheloniidae)

• 1.1. An intermediate morphotype between Eretmochelys imbricata and Caretta caretta was studied in Praia do Forte, Bahia, Brazil.
• 2.2. Three enzymatic systems were successfully analyzed: SOD, lactate dehydrogenase (LDH) and esterase (EST). Isoelectric focusing of total soluble proteins of muscle and transferrin were shown.
• 3.3. Esterase exhibited nine phenotype patterns, seven in C. caretta and one in the others morphotypes. SOD phenotypes were identical in the three morphotypes. Lactate dehydrogenase and transferrins were characteristic for each species.
• 4.4. Jaccard's measure of similarity was calculated and a phenogram with the three morphotypes were constructed using isoelectric focusing of total soluble proteins.

     

A carotenoid-pheophytin-protein complex from the digestive juice of silkworm (Bombyx mori L.)

• 1.1. A soluble carotenoid-pheophytin-protein complex was purified from the digestive juice of fifth instar silkworm larvae raised on mulberry leaves.
• 2.2. The pigment-protein complex showed absorption maxima at 276, 429, 453, 481 and 670 nm. Major pigment components were identified as α-carotene, pheophytin a and b.
• 3.3. This complex has an acidic protein component having an isoelectric point of 4.6. The molecular weight was estimated to be 68,000 with four identical subunits.

     

Superoxide dismutase,reduced glutathione and TBA-reactive products in erythrocytes of patients with multiple sclerosis

• 1.1. Superoxide dismutase, reduced glutathione and lipid peroxides levels were determined in the erythrocytes of multiple sclerosis patients.
• 2.2. Superoxide dismutase activity and the malonyldialdehyde production rate were found to be significantly enhanced.
• 3.3. The isoelectric focusing pattern of Superoxide dismutase from multiple sclerosis and normal subjects erythrocytes was substantially overlapping.
• 4.4. Our results indicate the occurrence of a higher susceptibility of multiple sclerosis erythrocytes to lipid peroxidation.

     

A novel inhibitor of glycogen phosphorylase from crayfish hepatopancreas

• 1.1. A novel glycogen phosphorylase inhibitor was partially purified from crayfish hepatopancreas.
• 2.2. The inhibitor was found only in two species of crayfish examined, and not in lobster, fresh and salt water clams, mussels or cockroaches.
• 3.3. The inhibitor is a small protein (M_r = 23,000) which did not show proteolytic activity.
• 4.4. Preliminary kinetic analysis of the inhibitory mechanism indicated that it bound to both glycogen and the glycogen phosphorylase protein.
• 5.5. Inhibitor binding to glycogen resulted in a competitive inhibition pattern with respect to glycogen phosphorylase (inhibition constant of ca 10 μg/ml).
• 6.6. The inhibitor also bound glycogen phosphorylase directly with a binding coefficient of 100 μg/ml resulting in a partially non-competitive inhibition pattern with respect to phosphate.

     

Alteration of the protein composition of bothrops jararaca venom and venom gland by isoproterenol treatment

• 1.1. The protein composition of Bothrops jararaca venom and venom gland was analyzed through SDS-PAGE, after isoproterenol (IPR) treatment.
• 2.2. Some proteins (47, 48, 57 and 72 kDa) were detected in the gland homogenate from the control but not from the IPR-treated samples.
• 3.3. Three proteins (26.5, 44.5 and 53 kDa) were detected in the venom gland from IPR-treated snakes but not from the venom gland from the control.
• 4.4. In the venom samples proteins of 41 and 74 kDa were detected only in the IPR treated samples, while proteins of 17 and 28 kDa were detected only in the control.
• 5.5. The biological activity of the venom did not change with IPR treatment.

     

Biochemical evidences for two sibling species of genus Myliobatis (chondrichthyes: myliobatidae) in South Brazil

• 1.1. Two morphotypes of Myliobatis from the demersal fishery off the Rio Grande (Brazil) were studied.
• 2.2. Thirty-two alleles were detected and resolved by 27 loci.
• 3.3. Nei's measure of genetic identity was 0.8306 and Thorpe's similarity was 0.6990. Mean heterozygosities observed were 0.1327 for the “DE” morphotype and 0.0409 for the “DL” morphotype.
• 4.4. Seven loci were fixed differently in the two taxa studied. This indicates the existence of a barrier to gene-flow between them, showing that both morphotypes belong to different species.
• 5.5. Jaccard's measure of similarity was calculated and a phenogram with the two morphotypes and M. freminvillii was constructed using isoelectric focusing of total soluble proteins. This showed a higher similarity between the two morphotypes of Myliobatis than M. freminvillii.