Starch Gel Zone Electrophoresis Research Articles

Biochemical variation and shell shape in populations of the fresh-water snail Lymnaea peregra (Mollusca, Gastropoda, Pulmonata) from south-west Ireland

Lymnaea peregra was collected at sites, predominantly from the Caha plateau, in south-west Ireland. These included sites investigated by previous workers and found to contain populations with a distinctive pattern of variation in shell-shape. Normally spired populations and non-spired (involuta, sub-involuta) could occur in adjacent loughs in the same watershed. The biochemical variation in some of these populations was investigated using horizontal starch-gel zone electrophoresis. Fourteen biochemical loci were investigated. Although circumscribed by difficulties in collecting and low sample numbers, the investigation showed that the pattern of variation in shell shape was still present and was not correlated or associated with the biochemical variation. While this did not support hypotheses of geographic race formation or incipient speciation, it was in agreement with the current view of variation and speciation in fresh-water pulmonates.

Immunological and physicochemical properties of a highly purified allergen from Dirofilaria immitis.

Allergen in crude extract of Dirofilaria immitis was purified and separated from IgG-inducing antigens by a combination of DEAE-Sephadex A-50 chromatography, Sephadex G-200 gel filtration and starch gel zone electrophoresis. The purified preparation was proved to be one protein band by sodium dodecyl sulfate polyacrylamide gel (SDS-gel) electrophoresis and one precipitin arc by immunodiffusion. The molecular weight of the purified allergen was estimated to be approximately 20,000 by gel filtration and 15,000 by SDS-gel electrophoresis. The carbohydrate content of the preparation was apparently low, about 2%. The allergen was positively charged, and its determinant group was protein in nature. It was resistant to tryptic, pepsic and chymotryptic digestion, periodate oxidation and DNase and RNase digestion but very sensitive to pronase digestion. Allergen was inclined to aggregate each other in the buffered solution. It was also very resistant to vibration, heat (80 degrees C for 1 h) and acid (pH 2.5) and alkali (pH 11.0) treatments. Rats as well as mice immunized with allergen developed only a reaginic antibody and no hemagglutination antibody.

Genetische Untersuchungen bei Picea abies mit Hilfe der Isoenzym-Identifizierung

Using techniques of starch gel zone electrophoresis, a considerable variation among esterase and leucine aminopeptidase isoenzyme patterns was found in the endosperm of dormant seeds of Norway spruce (Picea abies). Since the so-called primary endosperm of conifers is only a further developmental stage of the haploid female gametophyte, simple Mendelian segregations can be determined in seeds of individual open-pollinated trees. It was therefore possible to identify some esterase and leucine aminopeptidase loci only with regard to phenotypic frequency distributions without difficult crossing procedures.

Proteins in scorpion venoms toxic to mammals and insects.

Abstract The crude venoms of the scorpions Androctonus aeneas aeneas , Androctonus amoreuxi , Androctonus mauretanicus mauretanicus , Buthus occitanus pairs , Buthus occitanus tunetanus and Leiurus quinquestriatus were submitted to starch gel zone electrophoresis. The eluates of gel sections were assayed for mice lethality, contraction-paralysis of fly larvae and larvae lethality. It has been found that: (1) All the venoms contain discrete components showing a specific toxicity to mice and to fly larvae. (2) Three of the venoms contain more than one larvae contraction-paralysis component. (3) At pH 8·6, most of the mice toxic components have a cathodic mobility whereas the majority of larvae toxic components migrate towards the anode. (4) All the larvae contraction-paralysis components are strongly lethal to fly larvae and are inactivated by trypsin, thus demonstrating their protein nature. (5) The venoms of A. a. aeneas and B. o. tunetanus contain factions of fast anodic mobility which are lethal to fly larvae but lack the larvae contraction-paralysis and the mice lethal activities. It is concluded that the presence of a variety of toxic proteins specifically active on different animals is a general property of scorpion venoms.

Molecular differentiation of several enzymes in early human embryos

AbstractHomogenates of two human embryos, one each at horizons 16 and 20, were subjected to zone electrophoresis in starch and polyacrylamide gels. The gels were stained to reveal multiple molecular forms of acid and alkaline phosphatase, nonspecific esterase, and lactate, malate, and glucose‐6‐phosphate dehydrogenase. In general there were qualitative and quantitative increases in enzymatic activity with increasing gestational age. The enzymatic ontogeny observed was compared to that previously found prenatally in rats.

Electrophoretic protein patterns and esterase zymograms in ovaries and mature eggs of Bombyx mori in relation to diapause

Starch-gel zone electrophoresis was performed on proteins of pharate adult and emerged adult ovaries, and in mature eggs in adults, in order to ascertain the mode of action of the diapause hormone on protein metabolism in the silkworm, Bombyx mori. Many protein bands were displayed characteristically by the developing ovaries and mature eggs of the silkworm races used. Among these bands, only one was affected by the diapause hormone. That is, the protein band is intensive in ovaries and mature eggs of the non-diapause type, but weak in those of the diapause type. This band was named band S, migrating positively with a mobility similar to albumin. Non-specific esterase activities were also tested in pharate adult and emerged adult ovaries and in mature eggs in adults. Esterase bands were detected individually for each species used. Among these bands, one esterase band was very active in ovaries and mature eggs of the non-diapause type but weak in those of the diapause type or those inactivated by the diapause hormone. This esterase was named esterase A. A comparison was carried out between protein band S and esterase A as regards progression with time during pharate adult development, both with response to the suboesophageal ganglion and diapause hormone extracts and with electrophoretic mobility. Subsequently, it was suggested that band S possesses proteins including esterase A. This particular protein with esterase A activity is discussed from the standpoint of embryogeny.

A new toxic protein in the venom of the scorpion Androctonus australis Hector

By use of starch gel zone electrophoresis and Sephadex G-50 gel filtration a toxic protein with contraction-paralysis as well as lethal activity to fly larvae has been isolated from the venom of the scorpion Androctonus australis Hector. This larva-toxic protein is different from the mice-lethal neurotoxins of the same venom. The significance of this finding is discussed.

The biochemical genetics of rabbit erythrocyte esterases. Histochemical classification.

Esterases found in rabbit erythrocytes and platelets were separated by zone electrophoresis in starch gels. Activity tests in various substrates were used to classify the esterase bands into four major categories. The bands of each category were further subdivided into groups by the effects of various inhibiting and activating agents. These experiments showed that the genetic loci which control the enzymes of systems 1, 2 and 3 control isozymes that respond identically with the test reagents in their respective systems. Diisopropylfluorophosphate and eserine sulfate inhibition indicated that the active sites of systems 1 and 2 enzymes are different. This supported the conclusion that the esterases of systems 1 and 2 are the products of closely linked loci and not multiple alleles at a single locus. Carbonic anhydrases which migrated in the cathodal portion of the gel were identified by their sensitivity to acetazolamide and more intense staining with β- rather than α-naphthyl acetate as substrate.

Muscle Protein Polymorphism of Sablefish from the Eastern Pacific Ocean

Muscle proteins of 897 specimens of sablefish, collected during 1963–67 in the northeastern Pacific Ocean, ranging from Unalaska Island, Alaska, to Cape Blanco, Oregon, were analyzed by starch gel zone electrophoresis. Without exception, all specimens fitted into one of three phenotypic protein patterns resulting from two polymorphic proteins appearing singly or in pairs. Hereditary control by two codominant alleles, MuA and MuB, is postulated to explain the heterogeneity of the muscle protein patterns. Phenotypic distribution was shown to be independent of age and sex. The collection was arbitrarily divided into 10 areas, and the gene frequencies used as population parameters suggested homogeneity but did not provide positive evidence of heterogeneity in the sablefish inhabiting the waters of the eastern Pacific Ocean. The implications of these results are discussed in the light of previously reported racial and tagging studies.

Autosomal determination of erythrocyte glucose 6-phosphate dehydrogenase in domestic chickens and ring-necked pheasants

Erythrocyte glucose 6-phosphate dehydrogenase isozymes of domestic chickens, ring-necked pheasants, and their hybrids were studied, using the starch gel zone electrophoresis technique. In domestic chickens G6PD isozymes were represented by two fast-moving bands and an indistinct third band, whereas in ring-necked pheasants a slow-moving broad band which seemed to consist of two closely apposed G6PD isozymes was observed. The F1 hybrids showed three distinct bands combining the characteristic mobility pattern of the two parents, which seemed to indicate that both parental alleles are expressed in F1 hybrids. Since both male and female hybrids exhibited strikingly similar isozyme patterns representing both sire and dam, it was assumed that the genes controlling the production of G6PD in chicken and pheasant red blood cells are located on the autosomes.

NUCLEAR GENE CONTROL OF MITOCHONDRIAL MALIC DEHYDROGENASE IN MAIZE

The isozymes of malic dehydrogenase (MDH) of maize have been separated by starch gel zone electrophoresis. The MDH isozyme pattern was the same in different organs and developmental stages. The subcellular distribution of the different isozymes was established by means of differential centrifugation. The MDH isozymes were found to be differentially distributed in the cytoplasm, the mitochondria, and the glyoxysomes. The latter proved the only exception to the pattern constancy among organs insofar as they are present only in the scutella. Among several inbred maize strains examined, only variants of the mitochondrial isozymes were found. Crosses between strains with different isozyme patterns gave Mendelian inheritance: thus, these mitochondrial isozymes are under nuclear-gene control.

Contribution of Protein Electrophoresis to Rockfish (Scorpaenidae) Systematics

During 1965–67, multiple hemoglobins of 1905 specimens, comprising 2 species of Sebastolobus and 26 of Sebastodes; muscle proteins of 1677 specimens, comprising 1 species of Scorpaena, 1 of Scorpaenodes, 2 of Sebastolobus, 2 of Sebastes, and 24 of Sebastodes; and eye lens proteins of 431 specimens, covering 1 species of Sebastolobus and 19 of Sebastodes were examined by starch–gel zone electrophoresis. Scorpaena, Sebastolobus, and Sebastodes specimens were collected in the eastern Pacific Ocean (California to Gulf of Alaska), and Sebastes specimens were collected from the western Atlantic Ocean (Newfoundland). Electropherograms were independent of sex and size. Hemoglobin electropherograms were species specific, and where variant, nongenetically linked patterns appeared, evidence suggested existence of additional species, for example, S. diploproa (2), S. reedi (2), and S. aleutianus (3). Genetically linked polymorphism was detected in hemoglobin protein electropherograms of S. crameri and muscle protein electropherograms of S. elongatus. Species specificity broke down with muscle proteins in many species of Sebastodes resulting in the separation of the 27 species examined (from this and other studies) into four distinct subgroupings. Generically distinct patterns were prominently evident in the muscle proteins of Sebastodes, Sebastolobus, Scorpaenodes, and Scorpaena, but not in Sebastes. The order of difference between Sebastes and Sebastodes was no more than that inherent in the four subgroupings within the latter genus. Eye lens proteins were genus-specific only. Relative value of the three groups of proteins as diagnostic tools at various levels of organization and as characters in the phylogenetic analyses of scorpaenids is discussed.

The stepwise removal of histones from chicken erythrocyte nucleoprotein.

1. A fractionation of chicken erythrocyte histones was achieved simultaneously with their extraction from saline-washed nuclei by stepwise titrations to progressively lower pH values. 2. Different acids and dilute buffer solutions of comparable pH behaved similarly in stepwise extractions of histones. 3. The histone preparations so obtained were characterized by their amino acid composition and behaviour on zone electrophoresis in starch gels. 4. The fractionation by titration was quite sharp at appropriate pH ranges, and the histone fraction that is apparently unique to avian erythrocytes was obtained without contamination by other histone fractions. 5. Histones prepared by stepwise titration were fractionated further by cation-exchange and exclusion chromatography. The chromatographic behaviour and amino acid composition of the components permitted comparison with histones prepared by other methods. 6. Histone fraction IIb was resolved into its subfractions IIb(1) and IIb(2) by exclusion chromatography on Bio-Gel P-60. 7. Histone fractions III and IV, previously reported to be absent from chicken erythrocyte nuclei, were found in extracts made at pH1.

Separation of Pectolytic and Cellulolytic Enzymes in Culture Filtrates of Phytopathogenic Bacterial Species by Starch Gel Zone Electrophoresis

Separation of Pectolytic and Cellulolytic Enzymes in Culture Filtrates of Phytopathogenic Bacterial Species by Starch Gel Zone Electrophoresis

Characterization of the liver carboxylesterases of large domestic animals

1. 1. Aqueous extracts of porcine, bovine, ovine and equine livers were prepared and compared by zone electrophoresis in starch gel in continuous phosphate or borate buffer systems. 2. 2. The proteins and esterases were located in the starch gel by a variety of histochemical techniques. 3. 3. A partial characterization of the liver esterases from these species was carried out with various substrates and specific inhibitors. Species-specific electrophoretic patterns of esterase activity were observed. 4. 4. While most of the esterases of these species were identified as aliesterases or non-specific carboxylesterases (EC 3.1.1.1), acetylesterases (EC 3.1.1.6) were detected in porcine and ovine liver, and an active aryl esterase (EC 3.1.1.2) was observed in porcine liver.

Biochemical Changes During Sexual Development in Marchantia Polymorpha: I. Esterases

Extracts of uninduced thalli, induced thalli, stalks, antheridiophore and archegoniophore disks of Marchantia polymorpha were subjected to starch-gel zone electrophoresis. Developed gels were treated with appropriate reaction mixtures to detect sites of activity for 12 enzyme systems; only phosphatases, esterases, and peroxidases were observed. Although common sites of phosphatase, peroxidase, and esterase activity were detected in all tested extracts, additional sites of peroxidase and esterase activity were found in extracts from antheridiophore disks. The antheridia provided the additional esterases as determined by the electrophoresis of antheridial extracts and by a histochemical test for esterases in sections of antheridiophores.

Three new phenotypic systems in the blood of sheep.

SummaryThree new phenotypic systems detectable in the blood of sheep by means of the technique of zone electrophoresis in starch gels are described.One of these systems, designated EsA (A or arylesterase), involves a broad, smudge‐like zone of esterase which developed rapidly during staining and was observed in the serum of about ¼ of the animals sampled. The 2 phenotypes, referred to as EsA‐positive and EsA‐negative, are inherited as if controlled by a pair of alleles, EsA+ and EsA‐, with EsA+ being dominant to EsA‐. A quick tube test for the diagnosis of the 2 phenotypes is described. It is shown that the 2 phenotypes correspond respectively with the earlier described phenotypes Halon‐high and Halon‐low discovered by Lee in studies of the hydrolytic activity of ovine serum on di‐(2‐chloroethyl) aryl phosphates.A second system, designated CA, involves 2 electrophoretic forms, CA‐F and CA‐S, of erythrocytic carbonic anhydrase observed in 3 phenotypes CA‐F, CA‐FS and CA‐S. The 3 phenotypes are inherited as if controlled by co‐dominant autosomal alleles designated CAF and CAS. Allele CAS was at or near fixation in 6 of the 9 breeds surveyed in this study. This system has its analogue in the earlier described CA system of cattle.The third system, designated X (unknown) protein, involves 3 zones in electrophoresed red cell lysates which were either present together or absent together. The 2 phenotypes, designated X‐positive and X‐negative, are inherited as if controlled by a pair of alleles, X and x, with X being dominant to x. (This nomenclature is tentative and should be changed when the protein involved becomes identified.) The 2 phenotypes were observed in each of the 9 breeds in the population survey.

Characterization of rat liver and kidney esterases.

Vertical zone electrophoresis in starch gel was employed in conjunction with various histochemical techniques to characterize the liver and kidney esterases of male and female rats. Tissue-specific patterns were observed for each organ and a sex-dependent difference was noted on electrophoretic separation of the liver extracts. On the basis of substrate specificity and inhibitor sensitivity, both organs were observed to contain similar nonspecific aliesterases or carboxylesterases (EC 3.1.1.1). Inhibition studies showed the presence of two types of aliesterase in the liver and kidney extracts.

An Electrophoretic Survey of Esterases, Phosphatases, and Leucine Aminopeptidases in Mycelial Extracts of Species of Aspergillus

SUMMARYMycelial extracts of 32 strains representing 15 species of the genus Aspergillus were subjected to starch-gel zone electrophoresis to detect sites of leucine aminopeptidase, esterase, or phosphatase activity. Invariant and variant sites of esterase and phosphatase activity were found for 4 species. Esterase and phosphatase zymograms seemed to provide a biochemical means to distinguish species in this genus.

The Resolution of Four Lysine-rich Histones Derived from Calf Thymus

Abstract Histones rich in lysine were prepared by a procedure based on the direct extraction of calf thymus with aqueous trichloracetic acid. The protein obtained included what appeared to be degradation products, unless care was taken to isolate the histone rapidly (25 min) in the cold (0–3.5°) from thymus which had been frozen for transport to the laboratory. The protein preparations made according to this procedure appeared to be the same as other lysine-rich histone preparations described in the literature when examined by electrophoresis, chromatography, and amino acid analysis. These preparations seemed homogeneous in free boundary electrophoresis, and in zone electrophoresis in paper and starch gel. These preparations appeared homogeneous when examined by stepwise elution from carboxymethyl cellulose and when eluted from Amberlite IRC-50 by a gradient of barium acetate. Some resolution of these preparations was obtained by electrophoresis in polyacrylamide gel. This technique was especially powerful in detecting degradation products in the preparations. The most effective resolution of lysinerich histones, however, was obtained by chromatography on Amberlite IRC-50 with a gradient (7 to 14%) of guanidinium chloride in 0.1 m phosphate (pH 6.8). This method yielded three major fractions, one of which occasionally showed further partial resolution. Electrophoresis of the chromatographic fractions in polyacrylamide gel confirmed the presence of a total of four components in the lysine-rich histone prepared as noted above.