Characterization of an immunogenic fraction of Pasteurella multocida culture filtrates

1977 ◽  
Vol 23 (2) ◽  
pp. 197-201 ◽  
Author(s):  
Kunwar K. Srivastava ◽  
John W. Foster
Keyword(s):  
Culture Filtrate ◽  
Pasteurella Multocida ◽  
Gel Filtration ◽  
The Other ◽  
Rabbit Skin ◽  
Culture Filtrates ◽  
Homologous Challenge

An immunogenic fraction (IF) of Pasteurella multocida strain P-1059 was separated from culture filtrate by Sephadex gel filtration. Additional fractionation of IF with aqueous ether resulted in the glycoprotein-like preparation (GLP) while extraction with aqueous phenol provided the lipopolysaccharide-like preparation (LPP). The unextracted IF contained carbohydrate, protein, and lipid; the GLP contained carbohydrate and protein; and the LPP contained carbohydrate and lipid. The GLP was maximally protective for mice against homologous challenge, and was medially toxic in rabbit skin when compared to the other culture-filtrate preparations; the LPP was maximally toxic in rabbit skin, and was least protective for mice; and the unextracted IF was medially protective for mice, and was least toxic in rabbit skin.

Rapid Isolation of High Molecular Weight Urokinase from Native Human Urine

1982 ◽  
Vol 47 (03) ◽  
pp. 197-202 ◽  
Author(s):  
Kurt Huber ◽  
Johannes Kirchheimer ◽  
Bernd R Binder
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Human Urine ◽  
Gel Filtration ◽  
Chain Structure ◽  
The Other ◽  
Single Chain ◽  
Apparent Homogeneity ◽  
Sequential Adsorption

SummaryUrokinase (UK) could be purified to apparent homogeneity starting from crude urine by sequential adsorption and elution of the enzyme to gelatine-Sepharose and agmatine-Sepharose followed by gel filtration on Sephadex G-150. The purified product exhibited characteristics of the high molecular weight urokinase (HMW-UK) but did contain two distinct entities, one of which exhibited a two chain structure as reported for the HMW-UK while the other one exhibited an apparent single chain structure. The purification described is rapid and simple and results in an enzyme with probably no major alterations. Yields are high enough to obtain purified enzymes for characterization of UK from individual donors.

Experimental nephritis due to type specific streptococci. VI. Further characterization of type 12 nephrotoxin

1969 ◽  
Vol 15 (6) ◽  
pp. 555-561 ◽  
Author(s):  
P. W. Fardy ◽  
B. H. Matheson ◽  
R. W. Reed
Keyword(s):  
Active Material ◽  
Gel Filtration ◽  
Acute Glomerulonephritis ◽  
Acid Hydrolysate ◽  
Culture Filtrates ◽  
Experimental Nephritis ◽  
Chemical Studies ◽  
Group A ◽  
High Voltage Electrophoresis

Additional studies were undertaken on the nature of a nephrotoxic agent found in the culture filtrates of certain group A streptococci. A commercially available dehydrated medium proved satisfactory for the production of the active material. Gel filtration was used to divide polypeptide extracts, prepared from the dialyzable portion of culture filtrates, into two major fractions. One of these, representing the higher molecular weight components, contained most of the nephrotoxic activity as evidenced by the development of hypertension and acute glomerulonephritis in rabbits injected with this fraction.Physical and chemical studies indicated that the active fraction consisted of at least four polypeptide components separable by high voltage electrophoresis on paper. Automatic amino acid analysis of an acid hydrolysate of this fraction revealed 17 different amino acids. Carbohydrate was not detected by anthrone and orcinol tests.No relationship was established between this streptococcal nephrotoxic agent and other streptococcal constituents which have been implicated in acute glomerulonephritis.

Purification and Characterization of Two Proteins from Culture Filtrates of Mycobacterium tuberculosis H 37 Ra Strain

1970 ◽  
Vol 1 (2) ◽  
pp. 164-168
Author(s):  
Thomas M. Daniel ◽  
Lavenia E. Ferguson
Keyword(s):  
Molecular Weight ◽  
Mycobacterium Tuberculosis ◽  
Skin Testing ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Purification And Characterization ◽  
Culture Filtrates ◽  
Zone Electrophoresis

Two proteins have been purified from culture filtrates of Mycobacterium tuberculosis , H 37 Ra strain by a procedure combining gel filtration, diethylaminoethyl (DEAE)-cellulose chromatography, and zone electrophoresis. The two proteins are similar in molecular weight but differ slightly in charge. The faster migrating protein, designated a 1 , is not antigenic. The slower migrating protein, designated a 2 , is antigenic both with respect to antisera and as a skin-testing antigen.

scholarly journals THE PROTEINS IN UNHEATED CULTURE FILTRATES OF HUMAN TUBERCLE BACILLI

1948 ◽  
Vol 87 (3) ◽  
pp. 245-258 ◽  
Author(s):  
Janet R. McCarter ◽  
Ellen B. Bevilacqua
Keyword(s):  
Culture Filtrate ◽  
Guinea Pigs ◽  
The Other ◽  
Skin Tests ◽  
Sedimentation Constant ◽  
Culture Filtrates

1. Only two serologically different proteins were found in the unheated culture filtrates of both virulent and slightly virulent tubercle bacilli. One of them was the protein which had a sedimentation constant of 3.4 S, and the other was in filtrate fractions with a constant of 2 S. 2. That these proteins were distinct was demonstrated by three methods: quantitative precipitin and precipitin absorption tests with rabbit antisera, skin tests in guinea pigs actively sensitized with the culture filtrate fractions, and skin tests in passively sensitized guinea pigs. 3. A third antigen of unknown nature was found by means of the precipitin tests, but only in certain fractions from the virulent culture filtrate. 4. The protein with the constant of 3.4 S could not be demonstrated serologically in an O.T. made from the same culture filtrate as the unheated preparation from the virulent organism.

SOLUBILIZATION AND PARTIAL CHARACTERIZATION OF THYROID MEMBRANE TSH BINDING PROTEINS

1979 ◽  
Vol 92 (3) ◽  
pp. 512-521 ◽  
Author(s):  
B. Czarnocka ◽  
J. Nauman ◽  
G. Adler ◽  
W. Kiełczyński
Keyword(s):  
Binding Sites ◽  
Plasma Membranes ◽  
Binding Proteins ◽  
Gel Filtration ◽  
Soluble Proteins ◽  
The Other ◽  
High Affinity ◽  
Triton X ◽  
Maximal Capacity

ABSTRACT Crude plasma membranes obtained from bovine thyroids were found to possess one class of high affinity, low capacity binding sites for TSH with average association constant (Ka) of 1.301 × 109 m−1 and maximal capacity 8.76 × 10−10 m/mg of protein. Treatment of crude membranes fraction with 0.1 % Triton X-100 and the subsequent sonication in ultrasonic disintegrator resulted in solubilization of membranes proteins with mean recovery of 40.0 ± 6.2 %. Soluble proteins retained the property to bind [125I]TSH, but the binding of the hormone was decreased. The removal of the detergent from the solubilizate by gel filtration on Sephadex LH-20 increased the binding of TSH well above that demonstrated for crude thyroid membranes. The chromatography of soluble proteins on Ultrogel AcA-44 revealed the presence of two TSH binding proteins, one with the molecular weight (m.w.) above 130 000 daltons and the other with the m.w. approximately 30 000 daltons. The electrofocusing of solubilizate on Ampholine resulted in two protein peaks, one at pH 4.0–4.1 and the other at pH 4.4–4.6. The latter peak was shown to bind [125I]TSH specifically. The present results have confirmed the heterogeneous character of solubilized TSH receptor preparation and have shown that the hormone binding sites belong to acid proteins.

scholarly journals Partial characterization of the oligosaccharides of mouse thymocyte Thy-1 glycoprotein

1984 ◽  
Vol 221 (2) ◽  
pp. 379-392 ◽  
Author(s):  
S R Carlsson ◽  
T Stigbrand
Keyword(s):  
Gel Filtration ◽  
Glycosylation Site ◽  
The Other ◽  
Complex Type ◽  
Mouse Thymocyte ◽  
Lentil Lectin ◽  
High Mannose ◽  
Sepharose 4B ◽  
Oligosaccharide Structures

Four glycopeptides (I, IIA, IIB, III) with different oligosaccharide structures were isolated from purified mouse thymocyte Thy-1 glycoprotein. The glycoprotein was digested with Pronase, and the glycopeptide fraction was isolated by gel filtration and acetylated with [3H]acetic anhydride. The different glycan structures were separated by affinity chromatography on concanavalin A-Sepharose 4B and lentil lectin-Sepharose 4B. Size determinations of intact and exoglycosidase- and endoglycosidase-digested glycopeptides were performed by gel filtration on Bio-Gel P-6, calibrated with glycopeptides of known structure. On the basis of these experiments and on the behaviour of the glycopeptides on the lectin columns, the following structures of the oligosaccharide chains were proposed: I, triantennary ‘complex-type’ with terminal fucose; IIA, biantennary ‘complex-type’ without fucose; IIB, biantennary ‘complex-type’ with fucose; III, a mixture of ‘high-mannose’ chains containing either five or six mannose residues (approx. 50% of each). Amino acid analysis of the glycopeptides showed that the predominant oligosaccharide at glycosylation-site Asn-23 was of ‘high-mannose’ type, whereas the other two sites (Asn-75 and Asn-99) were glycosylated with ‘complex-type’ chains. Both these sites were shown to be variably glycosylated. The major glycans linked to Asn-75 were of structures I and IIB, whereas all three ‘complex-type’ chains were represented at Asn-99. The results presented explain the previously reported carbohydrate heterogeneity of thymocyte Thy-1 glycoprotein.

scholarly journals The xylanase system of Coniophora cerebella

1968 ◽  
Vol 108 (4) ◽  
pp. 571-576 ◽  
Author(s):  
N. J. King ◽  
D. B. Fuller
Keyword(s):  
Molecular Weight ◽  
Culture Filtrate ◽  
Uronic Acid ◽  
Enzyme System ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
The Other ◽  
Xylose Residue ◽  
Random Manner ◽  
Acidic Sugars

1. The culture filtrate of the fungus Coniophora cerebella grown on poplar 4-O-methylglucuronoxylan as carbon source and enzyme inducer contained an enzyme system that degraded the polysaccharide to xylose, acidic and neutral oligosaccharides and an enzyme-resistant polymer. Free uronic acid was not produced. 2. Cold ethanol fractionation of the culture filtrate yielded two active fractions, one of which had only xylanase (EC 3.2.1.8) and the other both xylanase and xylosidase (EC 3.2.1.37) activities. Further fractionation on DEAE-cellulose resolved the xylanase and xylosidase activities. 3. The xylanase degraded poplar 4-O-methylglucuronoxylan in an essentially random manner, producing oligosaccharides, but some xylose residues in the vicinity of uronic acid side groups were protected from hydrolysis, preventing a truly random attack. The xylosidase attacked the polysaccharide very slowly, releasing xylose, but the oligosaccharides produced by the action of the xylanase were much more susceptible to hydrolysis by the xylosidase. 4. The products of xylanase action were separated into neutral and acidic fractions. The neutral oligosaccharides were separated by chromatography on charcoal–Celite, and the major products were characterized as xylobiose, xylotriose, xylotetraose and xylopentaose. Some of the acidic sugars were branched, having the uronic acid residue attached to a xylose residue other than the terminal non-reducing one. 5. Gel filtration of various xylanase fractions gave values for the molecular weight of the enzyme from 34000 to 38000.

scholarly journals Characterization of human heterophil hemolysins induced by Schistosoma mansoni infection

1983 ◽  
Vol 78 (3) ◽  
pp. 257-267 ◽  
Author(s):  
Munir Chamone ◽  
L. Alves Oliveira ◽  
P. M. Z. Coelho ◽  
G. Gazzinelli ◽  
A. Oliveira Lima ◽  
...  
Keyword(s):  
Heat Treatment ◽  
Schistosoma Mansoni ◽  
Hemolytic Activity ◽  
Gel Filtration ◽  
The Other ◽  
Other Hand ◽  
Classic Pathway

Heterophil antibodies could be detected in sera from normal or from patient with chronic schistosomiasis. Their hemolytic activities depend on the integrity of the complement classic pathway. The heterophil antibodies from patient sera presented a higher specificity for Schistosoma mansoni antigen preparations than those detected in normal sera. Most of the hemolytic activity observed in normal sera can be destroyed at 56ºC for 4 min. On the other hand, about 80% of the sera from infected patients are partially or totally resistant to this heat-treatment. The hemolytic activities of sera were eluted from a gel filtration column in different fractions of the first peak.

scholarly journals Human small-intestinal β-galactosidases. Separation and characterization of one lactase and one hetero β-galactosidase

1969 ◽  
Vol 114 (2) ◽  
pp. 351-359 ◽  
Author(s):  
Nils-Georg Asp ◽  
Arne Dahlqvist ◽  
Otakar Koldovský
Keyword(s):  
Intestinal Mucosa ◽  
Gel Filtration ◽  
Competitive Inhibitor ◽  
The Other ◽  
Artificial Substrates ◽  
Small Intestinal Mucosa ◽  
Lactase Activity ◽  
Ph Optimum ◽  
Small Intestinal

1. Two β-galactosidases from human small-intestinal mucosa were separated by gel-filtration chromatography and the properties of the two enzymes were studied. Lactose and four hetero β-galactosides were used as substrates. 2. One of the enzymes was particle-bound and could be partially solubilized with papain. Of the substrates hydrolysed by this enzyme, lactose was hydrolysed most rapidly. This enzyme is thus essentially a disaccharidase and is named lactase. It is presumably identical with the ‘lactase 1’ described earlier. 3. The other enzyme was mainly soluble and hydrolysed all artificial substrates used, whereas no lactase activity could be detected. This enzyme has therefore been designated hetero β-galactosidase. 4. p-Chloromercuribenzoate (0·1mm) inhibited the hetero β-galactosidase completely but did not influence the activity of the lactase. Tris was a competitive inhibitor of both enzymes. 5. The residual lactase activity in the mucosa of lactose-intolerant patients may be exerted by a small amount of remaining lactase as such, or possibly by a third enzyme with a more acid pH optimum.

IMMUNOCHEMICAL CHARACTERIZATION OF PROTEINS IN CATTLE URINE AND THEIR COMPLEXES WITH SILICIC ACID

1978 ◽  
Vol 58 (3) ◽  
pp. 435-442
Author(s):  
C. B. BAILEY
Keyword(s):  
Urinary Tract ◽  
Sialic Acid ◽  
Silicic Acid ◽  
Serum Proteins ◽  
Gel Filtration ◽  
Urinary Calculi ◽  
The Other ◽  
Single Protein ◽  
Serum Components

Electrophoresis of total non-dialyzable solids (TNDS) from cattle urine showed the presence of components with mobilities covering the same range as noted for serum. Although most of the components had mobilities in the albumin to α-globulin region, they appeared to contain more glycoprotein than serum components with similar mobilities. Immunoelectrophoresis of TNDS against rabbit antibovine serum and rabbit anti-TNDS revealed the presence of four components giving reactions of identity with serum proteins and of one that originated from the urinary tract. Gel filtration of TNDS on Sephadex G-200 produced six poorly resolved peaks, four of which were shown by immunoelectrophoresis to contain the four serum components of TNDS. The fifth peak contained the urinary tract component. This component constituted about half the total weight of the proteinaceous constituents of TNDS and contained considerably more sialic acid than the other components. The substance represented by the sixth peak obtained by gel filtration was unidentified because it failed to elicit antibody formation in the rabbit. It was low in protein and sialic acid but had a much larger absorbance at 280 nm than the other constituents. Reaction of TNDS with polymerized silicic acid removed half the TNDS from solution by formation of an insoluble complex. One component was completely removed from solution and portions of others were partially removed by this procedure. It was concluded that in the process of formation of siliceous urinary calculi in cattle, precipitation of polymerized silicic acid is a generalized process that does not rely on the presence in urine of any single protein species.

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