Identification of Supernatant and Mitochondrial Isozymes of Malate Dehydrogenase on Electropherograms Applied to the Taxonomic Discrimination of Walleye (Stizostedion vitreum vitreum), Sauger (S. canadense), and Suspected Interspecific Hybrid Fishes

1973 ◽  
Vol 30 (7) ◽  
pp. 927-938 ◽  
Author(s):  
J. W. Clayton ◽  
R. E. K. Harris ◽  
D. N. Tretiak
Keyword(s):  
Malate Dehydrogenase ◽  
White Muscle ◽  
Heat Stability ◽  
Heart Tissue ◽  
Staining Procedure ◽  
Stizostedion Vitreum ◽  
Specific Staining ◽  
Heat Stable ◽  
Dye Reduction

Walleye (Stizostedion vitreum vitreum) and sauger (S. canadense) commonly yield four isozymes of malate dehydrogenase (MDH) in electropherograms of white muscle extracts from individual homozygous fish. The heat stability of each isozyme as well as reactivity with nicotinamide-adenine dinucleotide (NAD) and NAD analogues have been investigated by measuring the density of each isozyme directly on the electropherograms, after the usual specific staining procedure that links malate oxidation to tetrazolium dye reduction. At least two kinds of supernatant and one kind of mitochondrial MDH in individual fish of each species was demonstrated.Relative abundance of the various isozymes varies with the tissue of origin. In liver, only one heat stable isozyme is observed in both species, and it is likely constituted mainly of supernatant MDH. In heart tissue four isozymes are observed and the liver supernatant form also predominates. In white muscle the two kinds of supernatant MDH appear to be synthesized in comparable amounts and, including the mitochondrial MDH, four isozymes of nearly equivalent catalytic activity are produced. In walleye the three kinds of MDH homozygotes all showed similar enzymatic properties of their corresponding isozymes.In contrast to the polymorphic nature of walleye MDH isozymes the closely related sauger are monomorphic for MDH, and a number of analogous isozymes have identical electrophoretic mobility in the two species. More significantly, the major mitochondrial MDH isozymes have different mobility in each species. This fact is suggested as a taxonomic criterion to distinguish the two species. Some very rare fishes, evidently interspecific hybrids, produced three mitochondrial MDH isozymes. It was also possible to hybridize walleye and sauger MDH isozymes in vitro to produce phenotypes indistinguishable from presumed wild hybrid fishes.

scholarly journals Preliminary characterization of a heat-stable protein from rat adipose tissue whose phosphorylation is stimulated by insulin

1982 ◽  
Vol 204 (3) ◽  
pp. 817-824 ◽  
Author(s):  
P J Blackshear ◽  
R A Nemenoff ◽  
J Avruch
Keyword(s):  
Adipose Tissue ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Gel Filtration ◽  
Heat Stability ◽  
Intact Cells ◽  
Heat Stable ◽  
Heat Stable Protein ◽  
Rat Adipose Tissue

Exposure of 32P-labelled isolated rat adipocytes or epididymal fat-pads to insulin resulted in an increase in the phosphorylation of a heat-stable acid-soluble protein of Mr 22 000. The phosphorylation of this protein was unaffected by isoprenaline (isoproterenol) in intact cells, nor was its phosphorylation catalysed by exposure in vitro to the cyclic AMP-dependent protein kinase or smooth-muscle myosin light-chain kinase. The properties of the Mr-22 000 protein include: heat-stability; solubility in 1% trichloroacetic acid; pI 4.9; elution at apparent Mr 37 500 on gel filtration; and it contains both phosphoserine and phosphothreonine. It can be distinguished from the heat-stable phosphatase inhibitor 1 of adipose tissue (inhibitor 1A) and the phosphorylated form of adipose-tissue myosin light chain by several criteria. Its identity, and the possible functional significance of the insulin-stimulated phosphorylation, remain problems for future study.

scholarly journals Malate dehydrogenases in phototrophic purple bacteria. Thermal stability, amino acid composition and immunological properties

1988 ◽  
Vol 252 (2) ◽  
pp. 595-600 ◽  
Author(s):  
M A Tayeh ◽  
M T Madigan
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Malate Dehydrogenase ◽  
Acid Composition ◽  
Rhodospirillum Rubrum ◽  
Rhodobacter Capsulatus ◽  
Purple Bacteria ◽  
Heat Stability ◽  
Heat Stable ◽  
Rhodomicrobium Vannielii

Purified malate dehydrogenases from four species of non-sulphur purple phototrophic bacteria were examined for their heat-stability, amino acid composition and antigenic relationships. Malate dehydrogenase from Rhodospirillum rubrum, Rhodobacter capsulatus and Rhodomicrobium vannielii (which are all tetrameric proteins) had an unusually high glycine content, but the enzyme from Rhodocyclus purpureus (which is a dimer) did not. R. rubrum malate dehydrogenase was extremely heat-stable relative to the other enzymes, withstanding 65 degrees C for over 1 h with no loss of activity. By contrast, malate dehydrogenase from R. vannielii lost activity above 35 degrees C, and that from R. capsulatus above 40 degrees C. Amino acid compositional relatedness and immunological studies indicated that tetrameric phototrophic-bacterial malate dehydrogenases were highly related to one another, but only distantly related to the tetrameric enzyme from Bacillus. This suggests that, despite differences in their thermal properties, the tetrameric malate dehydrogenases of non-sulphur purple bacteria constitute a distinct biochemical class of this catalyst.

scholarly journals Multiplicity of clathrin light-chain-like polypeptides from developing pea (Pisum sativum L.) cotyledons

1992 ◽  
Vol 103 (4) ◽  
pp. 1127-1137 ◽  
Author(s):  
H.B. Lin ◽  
S.M. Harley ◽  
J.M. Butler ◽  
L. Beevers
Keyword(s):  
Pisum Sativum ◽  
Light Chain ◽  
Calcium Binding ◽  
Heat Stability ◽  
Coated Vesicles ◽  
Light Chains ◽  
Heavy Chains ◽  
Heat Stable ◽  
Pisum Sativum L

A comparative study has been made of clathrin-coated vesicles from developing pea (Pisum sativum L.) cotyledons and bovine brains in order to characterize the clathrin light chains from a plant system. Four polypeptides of 31 kDa, 40 kDa, 46 kDa and 50 kDa are considered as candidates for clathrin light chains in the developing pea cotyledons. The 31 kDa, 40 kDa, 46 kDa and 50 kDa polypeptides, together with the 190 kDa heavy chain, are dissociated as triskelions when coated vesicles of developing pea cotyledons are treated with 2 M urea. Partially purified 46 kDa and 50 kDa polypeptides have been demonstrated to bind to purified clathrin heavy chains. The 40 kDa, 46 kDa and 50 kDa polypeptides are sensitive to elastase. They are readily solubilized by neutralization of 10% trichloroacetic acid precipitates of clathrin. The 50 kDa polypeptide of plant clathrin-coated vesicles is heat-stable as are the light chains from bovine brains, while the heat stability of the 31 kDa, 40 kDa and 46 kDa polypeptides of plants is dependent on pH and ionic strength. The 40 kDa, 46 kDa and 50 kDa polypeptides bind calmodulin. The calcium binding properties of these polypeptides are ambiguous. The 40 kDa and 46 kDa polypeptides can be phosphorylated more extensively than the 31 kDa in vitro in the presence of polylysine, as can the smaller light chain of brains. The 50 kDa polypeptide can also be phosphorylated, even without the addition of polylysine. Unlike brain light chains, phosphorylation of the 31 kDa, 40 kDa, 46 kDa and 50 kDa polypeptides from peas is greatly reduced by N-ethylmaleimide (NEM). Our findings contrast with earlier reports of clathrin light chains of 30 and 38 kDa from zucchini and 57 and 60 kDa from carrots, respectively.

scholarly journals Guanidine and heat sensitivity of foot-and-mouth disease virus (FMDV) strains

1982 ◽  
Vol 89 (1) ◽  
pp. 129-138 ◽  
Author(s):  
P. F. Nettleton ◽  
Marilyn J. Davies ◽  
M. M. Rweyemamu
Keyword(s):  
Guanidine Hydrochloride ◽  
Foot And Mouth Disease ◽  
Heat Stability ◽  
Heat Inactivation ◽  
Heat Sensitivity ◽  
Fmd Virus ◽  
Heat Stable ◽  
Mouth Disease Virus ◽  
Stable Characteristic

SUMMARYA study of the ability of 49 strains of FMD virus to replicate in BHK-21 monolayer cells maintained under a standard agar overlay containing 5·2 mM guanidine hydrochloride and to withstand heat inactivation at 54°C for 1 h showed that strains belonging to serotypes C, O and Asia 1 were generally more resistant to guanidine and heat stable than the SAT 1, 2 and 3 serotypes. The type A viruses as a whole occupied an intermediate position between these two groups.In vitro passage in BHK-21 cells influenced the guanidine sensitivity of 3(O, C and SAT 3) of the 7 FMD serotypes suggesting that this is not a stable genetic marker. Heat stability of the FMD viruses, however, did not change on passage, suggesting that this is a stable characteristic inherent in any homogeneous FMD virus population.

scholarly journals C-Terminal Deletions Can Suppress Temperature-Sensitive Mutations and Change Dominance in the Phage Mu Repressor

Genetics ◽  
1996 ◽  
Vol 142 (3) ◽  
pp. 661-672 ◽  
Author(s):  
Jodi L Vogel ◽  
Vincent Geuskens ◽  
Lucie Desmet ◽  
N Patrick Higgins ◽  
Ariane Toussaint
Keyword(s):  
Binding Activity ◽  
Temperature Sensitive ◽  
Dna Binding Activity ◽  
Heat Stable ◽  
C Terminus ◽  
Acid Domain ◽  
Mutant Forms ◽  
Amino Acid Domain

Abstract Mutations in an N-terminal 70-amino acid domain of bacteriophage Mu's repressor cause temperature-sensitive DNA-binding activity. Surprisingly, amber mutations can conditionally correct the heat-sensitive defect in three mutant forms of the repressor gene, cts25 (D43-G), cts62 (R47-Q and cts71 (M28-I), and in the appropriate bacterial host produce a heat-stable Sts phenotype (for survival of temperature shifts). Sts repressor mutants are heat sensitive when in supE or supF hosts and heat resistant when in Sup° hosts. Mutants with an Sts phenotype have amber mutations at one of three codons, Q179, Q187, or Q190. The Sts phenotype relates to the repressor size: in Sup° hosts sts repressors are shorter by seven, 10, or 18 amino acids compared to repressors in supE or supF hosts. The truncated form of the sts62-1 repressor, which lacks 18 residues (Q179–V196), binds Mu operator DNA more stably at 42° in vitro compared to its full-length counterpart (cts62 repressor). In addition to influencing temperature sensitivity, the C-terminus appears to control the susceptibility to in vivo Clp proteolysis by influencing the multimeric structure of repressor.

scholarly journals Recapitulating Cardiac Structure and Function In Vitro from Simple to Complex Engineering

Micromachines ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 386
Author(s):  
Ana Santos ◽  
Yongjun Jang ◽  
Inwoo Son ◽  
Jongseong Kim ◽  
Yongdoo Park
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Computational Modeling ◽  
Cardiac Tissue ◽  
Cardiac Development ◽  
Heart Tissue ◽  
Cardiac Tissue Engineering ◽  
Cardiac Cells

Cardiac tissue engineering aims to generate in vivo-like functional tissue for the study of cardiac development, homeostasis, and regeneration. Since the heart is composed of various types of cells and extracellular matrix with a specific microenvironment, the fabrication of cardiac tissue in vitro requires integrating technologies of cardiac cells, biomaterials, fabrication, and computational modeling to model the complexity of heart tissue. Here, we review the recent progress of engineering techniques from simple to complex for fabricating matured cardiac tissue in vitro. Advancements in cardiomyocytes, extracellular matrix, geometry, and computational modeling will be discussed based on a technology perspective and their use for preparation of functional cardiac tissue. Since the heart is a very complex system at multiscale levels, an understanding of each technique and their interactions would be highly beneficial to the development of a fully functional heart in cardiac tissue engineering.

scholarly journals In vivo transcriptome analysis provides insights into host-dependent expression of virulence factors by Yersinia entomophaga MH96, during infection of Galleria mellonella

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Amber R Paulson ◽  
Maureen O’Callaghan ◽  
Xue-Xian Zhang ◽  
Paul B Rainey ◽  
Mark R H Hurst
Keyword(s):  
Galleria Mellonella ◽  
Functional Enrichment ◽  
Natural Environments ◽  
Insecticidal Toxin ◽  
Heat Stable ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Cell Densities

Abstract The function of microbes can be inferred from knowledge of genes specifically expressed in natural environments. Here, we report the in vivo transcriptome of the entomopathogenic bacterium Yersinia entomophaga MH96, captured during initial, septicemic, and pre-cadaveric stages of intrahemocoelic infection in Galleria mellonella. A total of 1285 genes were significantly upregulated by MH96 during infection; 829 genes responded to in vivo conditions during at least one stage of infection, 289 responded during two stages of infection, and 167 transcripts responded throughout all three stages of infection compared to in vitro conditions at equivalent cell densities. Genes upregulated during the earliest infection stage included components of the insecticidal toxin complex Yen-TC (chi1, chi2, and yenC1), genes for rearrangement hotspot element containing protein yenC3, cytolethal distending toxin cdtAB, and vegetative insecticidal toxin vip2. Genes more highly expressed throughout the infection cycle included the putative heat-stable enterotoxin yenT and three adhesins (usher-chaperone fimbria, filamentous hemagglutinin, and an AidA-like secreted adhesin). Clustering and functional enrichment of gene expression data also revealed expression of genes encoding type III and VI secretion system-associated effectors. Together these data provide insight into the pathobiology of MH96 and serve as an important resource supporting efforts to identify novel insecticidal agents.

scholarly journals A GENETICALLY DISTINCT FORM OF CYCLIC AMP PHOSPHODIESTERASE ASSOCIATED WITH CHROMOMERE 3D4 IN DROSOPHILA MELANOGASTER

Genetics ◽  
1979 ◽  
Vol 91 (3) ◽  
pp. 521-535
Author(s):  
John A Kiger ◽  
Eric Golanty
Keyword(s):  
Drosophila Melanogaster ◽  
Cyclic Amp ◽  
Structural Gene ◽  
Regulatory Gene ◽  
Heat Stability ◽  
Normal Female ◽  
Dependent Manner ◽  
Cyclic Amp Phosphodiesterase ◽  
Heat Stable ◽  
Camp Levels

ABSTRACT Two cyclic AMP phosphodiesterase enzymes (E.C.3.1.4.17) are present in homogenates of adult Drosophila melanogaster. The two enzymes differ from one another in heat stability, affinity for Mg++, Ca++ activation and molecular weight. They do not differ markedly in their affinities for cyclic AMP, and both exhibit anomalous Michaelis-Menten kinetics. The more heatlabile enzyme is controlled in a dosage-dependent manner by chromomere 3D4 of the X chromosome and is absent in flies that are deficient for chromomere 3D4. Chromomere 3D4 is also necessary for the maintenance of normal cAMP levels, for male fertility, and for normal female fertility and oogenesis. The structural gene(s) for the more heat-stable enzyme is located outside of chromomeres 3C12-3D4. Whether 3D4 contains a structural gene, or a regulatory gene necessary for the presence of the labile enzyme, remains to be determined.

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