scholarly journals THE PREPARATION AND PHYSICOCHEMICAL CHARACTERIZATION OF THE SERUM PROTEIN COMPONENTS OF COMPLEMENT

1941 ◽  
Vol 74 (4) ◽  
pp. 297-308 ◽  
Author(s):  
L. Pillemer ◽  
E. E. Ecker ◽  
J. L. Oncley ◽  
E. J. Cohn
Keyword(s):  
Ionic Strength ◽  
Guinea Pig ◽  
Potassium Chloride ◽  
Electrophoretic Mobility ◽  
Phosphate Buffer ◽  
Physicochemical Characterization ◽  
Sedimentation Constant ◽  
Protein Components ◽  
Pig Serum

1. Methods for the separation from guinea pig serum in highly purified form of three of the components of complement are described. These substances are the so called mid-piece, end-piece, and 4th component. 2. Mid-piece has been separated as a euglobulin, with an electrophoretic mobility of 2.9 x 10–5 in phosphate buffer of ionic strength 0.2 at pH 7.7, and with a sedimentation constant of 6.4 x 10–13 in potassium chloride of ionic strength 0.2. 3. End-piece and 4th component were found together in a euglobulin fraction of serum which contained 10.3 per cent carbohydrate and had an electrophoretic mobility of 4.2 x 10–5 in phosphate buffer of ionic strength 0.2 at pH 7.7.

scholarly journals THE PROPERTIES OF MAMMALIAN STRIATED MYOFIBRILS ISOLATED BY AN ENZYMATIC METHOD

1950 ◽  
Vol 91 (6) ◽  
pp. 655-664 ◽  
Author(s):  
Armin F. Schick ◽  
George M. Hass
Keyword(s):  
Ionic Strength ◽  
Potassium Chloride ◽  
Phosphate Buffer ◽  
Phosphate Buffer Solution ◽  
Potassium Phosphate ◽  
Microscopic Structure ◽  
Buffer Solution ◽  
Buffer Solutions ◽  
Alkaline Side ◽  
Fresh Frozen

A new method for the isolation of large numbers of individual myofibrils from fresh mammalian skeletal and cardiac muscle has been described. Purification of isolated myofibrils was accomplished by differential centrifugation of fresh frozen sections of muscle which had been mechanically agitated after exposure for 30 to 45 minutes at 0°C. to the action of a dilute solution of trypsin in a phosphate buffer solution with a pH of 7.0 and an ionic strength of 0.25. Isolated skeletal myofibrils of the rabbit and man have similar constant solubility properties. They dissolve in an aqueous mixture of 0.5 N potassium chloride and 0.03 N sodium bicarbonate, giving viscous solutions which exhibit conspicuous birefringence of flow. They are soluble in buffer solutions (ionic strength 0.15) on the acid side of pH 4 and alkaline side of pH 10. If the ionic strength of potassium phosphate buffer solutions is increased to 0.5 or if the ionic strength of phosphate-borate buffer solutions is increased to a similar value by addition of potassium chloride, the isolated myofibrils become soluble at neutrality. Hence, it is possible, first to isolate the myofibrils and then dissolve them without deviating appreciably from physiologic ranges of pH. The extent to which myofibrils are modified by the conditions imposed by the method of isolation is unknown. There is no significant change in microscopic structure or optical birefringence. Furthermore, there is retention of a form of physiological reactivity, for when the isolated skeletal myofibrils are immersed in solutions of adenosinetriphosphate, they promptly and irreversibly change from elongated fibrils with distinct structural detail into dense spherical masses without recognizable microscopic structure.

Purification and characterization of guinea pig serum aminoacylproline hydrolase (aminopeptidase P)

1992 ◽  
Vol 1119 (2) ◽  
pp. 140-147 ◽  
Author(s):  
James W. Ryan ◽  
Fernando Valido ◽  
Pierre Berryer ◽  
Alfred Y.K. Chung ◽  
James E. Ripka
Keyword(s):  
Guinea Pig ◽  
Purification And Characterization ◽  
Aminopeptidase P ◽  
Pig Serum

scholarly journals Effective intercalation of zein into Na-montmorillonite: role of the protein components and use of the developed biointerfaces

2016 ◽  
Vol 7 ◽  
pp. 1772-1782 ◽  
Author(s):  
Ana C S Alcântara ◽  
Margarita Darder ◽  
Pilar Aranda ◽  
Eduardo Ruiz-Hitzky
Keyword(s):  
Water Solution ◽  
Physicochemical Characterization ◽  
Layered Silicate ◽  
Aqueous Dispersion ◽  
Underlying Mechanism ◽  
Ethanol Medium ◽  
Major Storage Protein ◽  
Protein Components

Biohybrid materials based on the intercalation of zein, the major storage protein in corn, into sodium-exchanged montmorillonite were prepared following two synthesis strategies. The first one made use of zein dissolved in 80% (v/v) ethanol/water solution, the usual solvent for this protein, while the second method is new and uses a sequential process that implies the previous separation of zein components in absolute ethanol. This treatment of zein with ethanol renders a soluble yellow phase and an agglomerate of insoluble components, which are able to intercalate the layered silicate when an aqueous dispersion of montmorillonite is added to the ethanol medium containing both phases. The diverse steps in this second route were investigated individually in order to understand the underlying mechanism that drives to the intercalation of this complex hydrophobic biomacromolecule into the hydrophilic interlayer space of sodium-exchanged montmorillonite. In addition to physicochemical characterization of the resulting materials, these biohybrid interfaces were also evaluated as biofillers in the preparation of diverse ecofriendly nanocomposites.

scholarly journals The identification and characterization of two separate carboxylesterases in guinea-pig serum

1983 ◽  
Vol 215 (1) ◽  
pp. 91-99 ◽  
Author(s):  
K Cain ◽  
E Reiner ◽  
D G Williams
Keyword(s):  
Guinea Pig ◽  
Sodium Dodecyl Sulphate ◽  
Enzyme Preparation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Selective Inhibitor ◽  
Nitrophenyl Phosphate ◽  
Phosphate Treatment ◽  
Pig Serum

The esterase activity of guinea-pig serum was investigated. A 3-fold purification was achieved by removing the serum albumin by Blue Sepharose CL-6B affinity chromatography. The partially purified enzyme preparation had carboxylesterase and cholinesterase activities of 1.0 and 0.22 mumol of substrate/min per mg of protein respectively. The esterases were labelled with [3H]di-isopropyl phosphorofluoridate (DiPF) and separated electrophoretically on sodium dodecyl sulphate/polyacrylamide gels. Two main labelled bands were detected: band I had Mr 80 000 and bound 18-19 pmol of [3H]DiPF/mg of protein, and band II had Mr 58 000 and bound 7 pmol of [3H]DiPF/mg of protein. Bis-p-nitrophenyl phosphate (a selective inhibitor of carboxylesterase) inhibited most of the labelling of bands I and II. The residual labelling (8%) of band I but not band II (4%) was removed by preincubation of partially purified enzyme preparation with neostigmine (a selective inhibitor of cholinesterase). Paraoxon totally prevented the [3H]DiPF labelling of the partially purified enzyme preparation. Isoelectrofocusing of [3H]DiPF-labelled and uninhibited partially purified enzyme preparation revealed that there were at least two separate carboxylesterases, which had pI3.9 and pI6.2, a cholinesterase enzyme (pI4.3) and an unidentified protein that reacts with [3H]DiPF and has a pI5.0. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of these enzymes showed that the carboxylesterase enzymes at pI3.9 and pI6.2 corresponded to the 80 000-Mr subunit (band I) and 58 000-Mr subunit (band II). The cholinesterase enzyme was also composed of 80 000-Mr subunits (i.e. the residual labelling in band I after bis-p-nitrophenyl phosphate treatment). The unidentified protein at pI5.0 corresponded to the residual labelling in band II (Mr 58 000), which was insensitive to neostigmine and bis-p-nitrophenyl phosphate. These studies show that the carboxylesterase activity of guinea-pig serum is the result of at least two separate and distinct enzymes.

scholarly journals Separation and Characterization of Highly Charged Polyelectrolytes Using Free-Solution Capillary Electrophoresis

Polymers ◽  
2018 ◽  
Vol 10 (12) ◽  
pp. 1331
Author(s):  
Isabelle Desvignes ◽  
Joseph Chamieh ◽  
Hervé Cottet
Keyword(s):  
Capillary Electrophoresis ◽  
Ionic Strength ◽  
Charge Density ◽  
Electrophoretic Mobility ◽  
Separation Mechanism ◽  
Free Solution ◽  
Counterion Condensation ◽  
Charge Densities ◽  
Effective Mobility

The characterization of statistical copolymers of various charge densities remains an important and challenging analytical issue. Indeed, the polyelectrolyte (PE) effective electrophoretic mobility tends to level off above a certain charge density, due to the occurrence of Manning counterion condensation. Surprisingly, we demonstrate in this work that it is possible to get highly resolutive separations of charged PE using free-solution capillary electrophoresis, even above the critical value predicted by the Manning counterion condensation theory. Full separation of nine statistical poly(acrylamide-co-2-acrylamido-2-methylpropanesulfonate) polymers of different charge densities varying between 3% and 100% was obtained by adjusting the ionic strength of the background electrolyte (BGE) in counter electroosmotic mode. Distributions of the chemical charge density could be obtained for the nine PE samples, showing a strong asymmetry of the distribution for the highest-charged PE. This asymmetry can be explained by the different reactivity ratios during the copolymerization. To shed more light on the separation mechanism, effective and apparent selectivities were determined by a systematic study and modeling of the electrophoretic mobility dependence according to the ionic strength. It is demonstrated that the increase in resolution with increasing BGE ionic strength is not only due to a closer matching of the electroosmotic flow magnitude with the PE electrophoretic effective mobility, but also to an increase of the dependence of the PE effective mobility according to the charge density.

Dissociation and Reassociation of Poliovirus II. Protein Components Obtained by Urea Treatment of the Virus Particle

1977 ◽  
Vol 32 (7-8) ◽  
pp. 632-636 ◽  
Author(s):  
Ursula Yamaguchi-Koll ◽  
K. J. Wiegers ◽  
R. Drzeniek
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Virus Particle ◽  
Gel Electrophoresis ◽  
Phosphate Buffer ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Urea Treatment ◽  
Primary Products ◽  
Protein Components

Abstract Dissociation of poliovirus by 9 ᴍ urea in 0.015 ᴍ NaCl at 25 °C resulted in the liberation of 35S RNA and of polypeptides sedimenting at 2S in sucrose gradients containing 9 ᴍ urea. However. a ribonucleopolypeptide (RNPP) complex sedimenting at 45S and oligomers of the viral polypeptides sedimenting at 7 -8S were found in addition to the monomers sedimenting at 2S when the urea concentration was lowered to 5 M after the dissociation procedure. Ribonuclease treatment prevents the appearance of the RNPP-complex. The amount of the RNPP-complex de­ creased, when the dissociation was performed at higher ionic strength. Under these conditions small amounts of empty capsids were detected. Polyacrylamide gel electrophoresis showed that the RNPP-complex contained the polypeptide VP1. The oligomers (7 -8S) contained the polypeptide VP3 and small amounts of VP2. The bulk of VP2 and some VP3 were found in the 2S position together with VP4. The molecular weight of the dissociation products in urea and phosphate buffer was determined by gel filtration to be about 30,000 for the monomeric polypeptides containing predominantly VP2 and about 70,000 for the oligomeric polypeptides containing predominantly VP3. Our results demonstrate that the oligomers and the RNPP-complex are not primary products obtained by dissociation of the virus particle by urea but are due to a reassociation of the poly­ peptides or of VP1 and RNA.

scholarly journals Serologic and Physicochemical Characterization of Donath-Landsteiner Antibodies from Six Patients

Blood ◽  
1963 ◽  
Vol 22 (5) ◽  
pp. 600-605 ◽  
Author(s):  
CARL F. HINZ
Keyword(s):  
Electrophoretic Mobility ◽  
Physicochemical Characterization ◽  
Chromatographic Behavior ◽  
Γ Globulins

Abstract Donath-Landsteiner antibodies from six patients were studied serologically and physicochemically. All had similar serologic properties including a requirement for complement in the cold phase. Five of the antibodies were associated with the 7S γ2-globulins, while the sixth was associated with 7S γ-globulins of slightly faster electrophoretic mobility and slightly different chromatographic behavior.

scholarly journals THE IMMUNOCHEMISTRY OF TOXINS AND TOXOIDS

1948 ◽  
Vol 88 (2) ◽  
pp. 205-221 ◽  
Author(s):  
Louis Pillemer ◽  
Ruth G. Wittler ◽  
Jean I. Burrell ◽  
D. B. Grossberg
Keyword(s):  
Ionic Strength ◽  
Electrophoretic Mobility ◽  
Isoelectric Point ◽  
Molecular Species ◽  
Protein Concentration ◽  
Rabbit Serum ◽  
Clostridium Tetani ◽  
Sedimentation Constant ◽  
Veronal Buffer ◽  
Ultracentrifugal Analysis

Methods for the purification and crystallization of tetanal toxin are described. The methods consist of the multiphase fractionation system involving methanol as the organic precipitating agent under controlled conditions of pH, ionic strength, protein concentration, and temperature. Crystalline tetanal toxin has an electrophoretic mobility of 2.8 x 10–5 in veronal buffer of 0.1 ionic strength at pH 8.6. The solubility of freshly prepared toxin is essentially constant. The isoelectric point is 5.1 ± 0.1. The crystalline toxin contains 1 per cent sulfur, traces of phosphorus, and gives the usual protein reactions. It does not contain carbohydrate. The crystalline toxin does not precipitate anti-Clostridium tetani rabbit serum. The final product contains between 3400 and 3600 Lf and about 6.6 x 107 M.L.D. per mg. N. Crystalline tetanal toxin is spontaneously converted to a flocculating atoxic dimer upon standing at 0°. This change is accompanied by the appearance of another molecular species as judged by constant solubility tests. Ultracentrifugal analysis of these fractions reveals that tetanal toxin has a sedimentation constant of 4.5 Svedberg units while the atoxic flocculating dimer sediments at 7 S.

Sign in / Sign up

Export Citation Format

Share Document