Studies on Molecular Weights of Two Peptide Hormones from the Urophysis of White Sucker (Catostomus commersoni)

1975 ◽  
Vol 53 (2) ◽  
pp. 242-247 ◽  
Author(s):  
Graham Moore ◽  
Anita Letter ◽  
Maria Tesanovic ◽  
Karl Lederis
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Hypotensive Activity ◽  
White Sucker ◽  
Urotensin Ii ◽  
Catostomus Commersoni ◽  
Molecular Weights ◽  
Long Acting

The molecular weights of two active principles extracted from the urophysis of the teleost fish Catostomus commersoni in 0.1 N HCl or in 0.25% acetic acid have been investigated by gel filtration chromatography and SDS–polyacrylamide gel electrophoresis. Two peptides with urotensin I (long-acting rat hypotensive) activity and two peptides with urotensin II (fish smooth muscle stimulating) activity were found by these procedures. The smaller of the two urotensin I peptides (molecular weight 1200–1700), designated urotensin IS, was shown to be a fragment of the larger peptide (molecular weight 2300–3000) which is produced by acid hydrolysis without loss of rat hypotensive activity. The two urotensin II peptides are suggested to represent either a monomer and a dimer or open and closed forms of a peptide.

scholarly journals Characterization of proteins from the cytoskeleton of Giardia lamblia

1983 ◽  
Vol 59 (1) ◽  
pp. 81-103 ◽  
Author(s):  
R. Crossley ◽  
D.V. Holberton
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulphate ◽  
Giardia Lamblia ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Molecular Size ◽  
Molecular Weights

Proteins from the axonemes and disc cytoskeleton of Giardia lamblia have been examined by sodium dodecyl sulphate/polyacrylamide gel electrophoresis. In addition to tubulin and the 30 X 10(3) molecular weight disc protein, at least 18 minor components copurify with the two major proteins in Triton-insoluble structures. The most prominent minor bands have the apparent molecular weights of 110 X 10(3), 95 X 10(3) and 81 X 10(3). Protein of 30 X 10(3) molecular weight accounts for about 20% of organelle protein on gels. In continuous 25 mM-Tris-glycine buffer it migrates mostly as a close-spaced doublet of polypeptides, which are here given the name giardins. Giardia tubulin and giardin have been purified by gel filtration chromatography in the presence of sodium dodecyl sulphate. Well-separated fractions were obtained that could be further characterized. Both proteins are heterogeneous when examined by isoelectric focusing. Five tubulin chains were detected by PAGE Blue 83 dye-binding after focusing in a broad-range ampholyte gel. Giardin is slightly less acidic than tubulin. On gels it splits into four major and four minor chains with isoelectric points in the pI range from 5.8 to 6.2. The amino acid composition of the giardin fraction has been determined, and compared to Giardia tubulin and a rat brain tubulin standard. Giardins are rich in helix-forming residues, particularly leucine. They have a low content of proline and glycine; therefore they may have extensive alpha-helical regions and be rod-shaped. As integral proteins of disc microribbons, giardins in vivo associate closely with tubulin. The properties of giardins indicate that in a number of respects - molecular size, charge, stoichiometry - their structural interaction with tubulin assemblies will be different from other tubulin-accessory protein copolymers studied in vitro.

Purification and properties of mercuric reductase from Yersinia enterocolitica 138A14

1993 ◽  
Vol 39 (2) ◽  
pp. 193-200 ◽  
Author(s):  
Mohamed Blaghen ◽  
Dominique J. M. Vidon ◽  
Mohamed Said El Kebbaj
Keyword(s):  
Molecular Weight ◽  
Yersinia Enterocolitica ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Mercury Resistance ◽  
Thiol Compounds ◽  
Mercuric Reductase ◽  
Purification And Properties ◽  
Layer Chromatography

A mercuric ion-reducing flavoprotein was purified from Yersinia enterocolitica 138A14 using dye matrix affinity chromatography. The purified enzyme had a characteristic absorption spectrum similar to those of flavin compounds, and FAD was detected as a part of the purified enzyme by thin-layer chromatography. Freshly purified preparations of the enzyme showed a single band on SDS polyacrylamide gel electrophoresis with a molecular weight of 70 000. The isolated enzyme had a molecular weight of about 200 000 as determined by gel filtration and disc gel electrophoresis. These results suggest an apparently trimeric structure of the enzyme. Dithiothreitol treatment disrupted the trimer into a dimeric structure of 140 000. Along with ageing, as well as limited proteolytic digestion, the enzyme evolved to give a dimeric molecule of 105 000 composed of two identical subunits of 52 000. The combination of the purified enzyme with HgCl2, or unexpectedly with merthiolate, oxidised the NADPH, which was followed spectrophotometrically. The Km for HgCl2 was dependent on the concentration of exogenous thiol compounds. A comparison of physical properties as well as kinetic characteristics indicated that the enzyme from Y. enterocolitica 138A14 is similar to mercuric reductases isolated from other mercury-resistant bacteria.Key words: Yersinia enterocolitica, mercury resistance, mercuric reductase.

scholarly journals Molecular weight and subunit size of fatty acid synthetase from rabbit mammary gland

1976 ◽  
Vol 159 (1) ◽  
pp. 181-184 ◽  
Author(s):  
N Paskin ◽  
R J Mayer
Keyword(s):  
Molecular Weight ◽  
Mammary Gland ◽  
Fatty Acid ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Fatty Acid Synthetase ◽  
Subunit Size

Fatty acid synthetase purified from the mammary gland of the rabbit has a mol. wt. of 968000 as determined by gel filtration. The enzyme gave one band, corresponding to a mol.wt. of approx. 35000, on polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate and phenylmethanesulphonyl fluoride.

II. Mitt.: Auflösung der Chromoproteidfraktion aus Rattenleber-Ribosomen mit Hilfe der Polyacrylamidgel-Elektrophorese

1970 ◽  
Vol 25 (1) ◽  
pp. 67-72 ◽  
Author(s):  
Wolfram Domschke ◽  
Jürgen G. Meyer-Bertenrath
Keyword(s):  
Gel Electrophoresis ◽  
Ribosomal Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
The Other ◽  
Blue Staining ◽  
Ribosomal Subunits ◽  
Molecular Weights ◽  
The One ◽  
Liver Ribosomes

After preparation of a coloured protein component containing iron from rat liver ribosomes 1 this fraction was submitted to detailed analysis by means of polyacrylamide gel electrophoresis. Thus it may be separated into one main and two secondary bands, which do not contain RNA detectable by methylene blue staining. The ferric content of all bands can be demonstrated by staining with 2,4-dinitroso-1,3-naphthalenediol 2. These bands are to be found in the large ribosomal subunits as well as in the small one in qualitative conformity, they differ, however, in their quantitative relations to each other depending on origin. LiCl-extraction as described for the preparation of ribosomal proteins causes dissociation of the chromoproteid fraction into six bands possessing lower molecular weights each than the original bands.The chromoproteids, localized in the large ribosomal subunits on the one hand and in the small subunits on the other hand were prepared differentiatedly by gel filtration. Results show the chromoproteid represented in the 57-S-subunit on a bigger scale than the nucleoproteid part, on the contrary, the 29-S-subunit is constructed of RNA-containing material preferably.

Formation of Soluble Complexes of Fibrinogen Related Material During Ancrod Infusion

1975 ◽  
Author(s):  
Caroline McKillop ◽  
W. Edgar ◽  
C. D. Forbes ◽  
C. R. M. Prentice
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Polyacrylamide Gel ◽  
Agarose Gel ◽  
Blood Samples ◽  
Related Material ◽  
Beta Alanine ◽  
Soluble Complexes

Seven pationts undergoing therapeutic defibrination by ancrod infusion were studied. Blood samples were obtained before treatment and after 6 and 24 hours ancrod infusion. Fibrinogen and its derivatives were precipitated with beta-alanine and separated by ΰ per cent agarose gel filtration. A range of soluble complexes were demonstrated after 6 hours infusion. Polyacrylamide gel electrophoresis in SDS showed that the soluble complexes were largely composed of units with molecular weight similar to a minimally degraded early Fragment X. Polyacrylamide gel electrophoresis in SDS and mercaptoethanol showed a marked loss of intact alphachain in the soluble complexes when compared with the uncomplexed material, suggesting that the soluble complexes had undergone preferential fibrinolytic digestion. It is suggested that, during ancrod therapy, FDP may be produced directly from soluble complexes rather than insoluble micro-thrombi as has been suggested previously.

scholarly journals Purification and subunit structure of legumin of Vicia faba L. (broad bean)

1974 ◽  
Vol 141 (2) ◽  
pp. 413-418 ◽  
Author(s):  
David J. Wright ◽  
Donald Boulter
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Broad Bean ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Exchange Chromatography ◽  
Polyacrylamide Gel ◽  
Subunit Structure ◽  
Isoelectric Precipitation ◽  
Molecular Weights

Zonal isoelectric precipitation was shown to be an effective method for the preparation of legumin which was homogeneous as judged by ultracentrifugation and polyacrylamide-gel electrophoresis. The subunit structure of legumin was investigated by preparative sodium dodecyl sulphate–polyacrylamide-gel electrophoresis and ion-exchange chromatography in urea. Five distinct subunits, of which two were acidic (α) and had a molecular weight of 37000, and three were basic (β) with molecular weights of 20100, 20900 and 23800, were identified. The α and β subunits were present in equimolar amounts in the legumin molecule and, in view of this and molecular-weight considerations, an α6β6 subunit model was proposed for legumin.

scholarly journals The fractionation, characterization, and subcellular localization of colony-stimulating activities released by the human monocyte-like cell line, GCT

Blood ◽  
1980 ◽  
Vol 56 (4) ◽  
pp. 717-727
Author(s):  
JF DiPersio ◽  
JK Brennan ◽  
MA Lichtman ◽  
CN Abboud ◽  
FH Kirkpatrick
Keyword(s):  
Molecular Weight ◽  
Cell Line ◽  
Gel Electrophoresis ◽  
Disulfide Bonds ◽  
Polyacrylamide Gel Electrophoresis ◽  
Human Monocyte ◽  
Colony Growth ◽  
Subcellular Fractionation ◽  
Molecular Weights ◽  
Cell Cytosol

GCT, a human monocyte-like cell line, has been shown to release biochemically distinct colony-stimulating activities (CSAs) for mouse and human marrows. These appear to be periodate-sensitive proteins with critical disulfide bonds. One, of molecular weight 145,000 daltons, stimulates macrophagic colony growth and is related to a 30,000-dalton molecule that also stimulates mouse growth. A 30,000-dalton CSA for human marrow can be separated from the 30,000-dalton mouse CSA by isoelectric focusing and gradient polyacrylamide gel electrophoresis. This distinction agrees with our previous finding of differential neutralization with anti-human urinary CSF antibody. The 30,000-dalton CSAs stimulate neutrophil, neutrophil-monocyte, and eosinophil colony growth in human marrow but only neutrophil and neutrophil-monocyte colonies in the mouse. Subcellular fractionation of GCT cells indicates that there are pools of preformed CSAs primarily associated with the cell cytosol that have similar apparent molecular weights to their secreted counterparts.

scholarly journals New Exfoliative Toxin Produced by a Plasmid-Carrying Strain of Staphylococcus hyicus

1999 ◽  
Vol 67 (8) ◽  
pp. 4014-4018 ◽  
Author(s):  
Hisaaki Sato ◽  
Takao Watanabe ◽  
Yasuko Murata ◽  
Ayumi Ohtake ◽  
Mayumi Nakamura ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
The Other ◽  
Exfoliative Toxin ◽  
Sds Page ◽  
Serotype B

ABSTRACT A new serotype of Staphylococcus hyicus exfoliative toxin (SHET), serotype B, was isolated from the culture filtrate of a plasmid-carrying strain of S. hyicus. The new SHET was purified by precipitation with 70% saturated ammonium sulfate, gel filtration on a Sephadex G-75 column, column chromatography on DEAE–Cellulofine A-500, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The new SHET caused exfoliation of the epidermis as determined by the so-called Nikolsky sign when inoculated into 1-day-old chickens. The new SHET was serologically different fromStaphylococcus aureus exfoliative toxins (ETs) (ETA, ETB, and ETC) and from the SHET from the plasmidless strain but showed the same molecular weight as the other serotypes of toxins on SDS-PAGE. It was thermolabile and lost its toxicity after being heated at 60°C for 30 min. We propose that the new SHET be designated SHETB and that the SHET produced by the plasmidless strain be designated SHETA.

Analysis of proteins in rabbit pulmonary surfactant using monoclonal antibodies

1986 ◽  
Vol 250 (3) ◽  
pp. C460-C467 ◽  
Author(s):  
R. J. King ◽  
H. M. Martin ◽  
J. B. Baseman ◽  
J. Morrison-Plummer
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
High Molecular Weight ◽  
Gel Electrophoresis ◽  
Pulmonary Surfactant ◽  
Polyacrylamide Gel Electrophoresis ◽  
Low Molecular Weight ◽  
Weight Group ◽  
Molecular Weights ◽  
Intermediate Group

We have used monoclonal antibodies developed against the apolipoproteins associated with pulmonary surfactant purified from rabbit lavage fluid to study the expression of epitopes common to these proteins. The pulmonary surfactant contained nearly 20 proteins, of which at least 10 were not derived from serum. Electrophoresis, with sulfhydryl reduction of these proteins indicated apparent molecular weights of approximately 155, 135, 125, and 115 X 10(3) (high-molecular-weight group); 80, 70, and 60 X 10(3) (intermediate group); and 18 through 10 X 10(3) (low-molecular-weight group). Two-dimensional polyacrylamide gel electrophoresis, in which the proteins were electrophoresed without reduction in the first dimension, but with sulfhydryl reduction in the second dimension, revealed that the 80, 70, and 60 X 10(3) proteins dissociated into proteins of nominal molecular weights of 40, 35, and 30 X 10(3), respectively. In contrast, the 125 and 115 X 10(3) proteins of the high-molecular-weight group contained a protein which could only be reduced to a minimum molecular weight of 55 to 60 X 10(3). Monoclonal antibodies generally were of three types: those that reacted strongly with the high-molecular-weight group and weakly with the intermediate group; those that reacted conversely; and those that reacted only with the low-molecular-weight group. Our results indicate that at least two different surfactant apolipoproteins, with differing minimum molecular weights in SDS-polyacrylamide gel electrophoresis, have common epitopes. Although these results cannot certify a physiological relationship between these proteins, they suggest that the intracellular synthesis or extracellular processing of surfactant apolipoproteins may be more complicated than predicted by the findings of previous experiments, perhaps involving the posttranslational assembly of one surfactant protein into oligomers which resist dissociation under the conditions used for the analyses.

Sign in / Sign up

Export Citation Format

Share Document