Identification of two core types in lipopolysaccharides ofActinobacillus pleuropneumoniaerepresenting serotypes 1 to 12

1996 ◽  
Vol 42 (8) ◽  
pp. 855-858 ◽  
Author(s):  
Mario Jacques ◽  
Stéphane Rioux ◽  
Sonia-Élaine Paradis ◽  
Caroline Bégin ◽  
Marcelo Gottschalk
Keyword(s):  
Western Blot ◽  
Salmonella Typhimurium ◽  
Gel Electrophoresis ◽  
Lipid A ◽  
Polyacrylamide Gel Electrophoresis ◽  
Actinobacillus Pleuropneumoniae ◽  
Type I ◽  
Type Ii ◽  
The Core ◽  
Core Lipid

Lipopolysaccharides (LPS) of Actinobacillus pleuropneumoniae were separated by Tricine–SDS–polyacrylamide gel electrophoresis, which has been shown to improve resolution of low-molecular-mass fast migrating bands. Strains representing the 12 serotypes of A. pleuropneumoniae can be divided in two groups according to the gel mobility of the core–lipid A region of their LPS. The first electromorphic core type (core type I), found in serotypes 1, 6, 9, and 11, had a migration slower than Salmonella typhimurium Ra LPS. The second electromorphic core type (core type II), found in the remaining serotypes (i.e., 2, 3, 4, 5, 7, 8, 10, and 12) had a migration similar to S. typhimurium Ra LPS. Furthermore, we observed that these two core types were antigenically different. Western blot analyses indicated that core – lipid A region of LPS from electromorphic core type I strains reacted when probed with serum from a pig experimentally infected with a core type I strain but not when probed with serum from a pig experimentally infected with a core type II strain. Conversely, core – lipid A region of LPS from electromorphic core type II strains reacted only when probed with serum from a pig experimentally infected with a core type II strain. Our results, based on both electrophoretic mobility and antigenicity, suggest the presence of two LPS core types in A. pleuropneumoniae.Key words: Actinobacillus pleuropneumoniae, lipopolysaccharides, core.

scholarly journals Interaction between proteoglycan subunit and type II collagen from bovine nasal cartilage, and the preferential binding of proteoglycan subunit to type I collagen

1976 ◽  
Vol 153 (2) ◽  
pp. 259-264 ◽  
Author(s):  
V Lee-Own ◽  
J C Anderson
Keyword(s):  
Gel Electrophoresis ◽  
Type I Collagen ◽  
Polyacrylamide Gel Electrophoresis ◽  
Type Ii Collagen ◽  
Type I ◽  
Type Ii ◽  
Nasal Cartilage ◽  
Preferential Binding ◽  
Calf Skin

We studied the interaction of proteoglycan subunit with both types I and II collagen. All three molecular species were isolated from the ox. Type II collagen, prepared from papain-digested bovine nasal cartilage, was characterized by gel electrophoresis, amino acid analysis and CM-cellulose chromatography. By comparison of type I collagen, prepared from papain-digested calf skin, with native calf skin acid-soluble tropocollagen, we concluded that the papain treatment left the collagen molecules intact. Interactions were carried out at 4 degrees C in 0.06 M-sodium acetate, pH 4.8, and the results were studied by two slightly different methods involving CM-cellulose chromatography and polyacrylamide-gel electrophoresis. It was demonstrated that proteoglycan subunit, from bovine nasal cartilage, bound to cartilage collagen. Competitive-interaction experiments showed that, in the presence of equal amounts of calf skin acid-soluble tropocollagen (type I) and bovine nasal cartilage collagen (type II), proteoglycan subunit bound preferentially to the type I collagen. We suggest from these results that, although not measured under physiological conditions, it is unlikely that the binding in vivo between type II collagen and proteoglycan is appreciably stronger than that between type I collagen and proteoglycan.

GCDFP-70 protein in cyst fluid identified as albumin and used to classify cysts in women with breast gross cystic disease

1991 ◽  
Vol 37 (4) ◽  
pp. 547-551 ◽  
Author(s):  
Milagros Balbín ◽  
Francisco Vizoso ◽  
Luis M Sánchez ◽  
Rafael Venta ◽  
Alvaro Ruibal ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Simple Procedure ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cyst Fluid ◽  
Type I ◽  
Type Ii ◽  
Cystic Disease ◽  
Functional Changes ◽  
Cyst Fluids

Abstract We used sodium dodecyl sulfate-polyacrylamide gel electrophoresis to study cyst fluids from women with breast gross cystic disease. The subjects could be classified into two categories according to the concentrations of protein GCDFP-70 in the cyst fluid: those with Type I cysts had a very low content of this protein; those with Type II cysts had very high concentrations. Analysis of the amino acid sequence of GCDFP-70 from both cyst fluid types confirmed that this protein is human plasma albumin. The average concentration of albumin found in Type I cyst fluids was 0.32 g/L and that corresponding to Type II was 10.16 g/L. Thus, albumin quantification from cyst fluids or analysis by either polyacrylamide or agarose gel electrophoresis provides a simple procedure for classifying these fluids, yielding results that correlate well with previous classifications based on other measurements such as sodium, potassium, and chloride concentrations. This albumin-based quantification method may improve the classification of breast cysts and might be useful in further studies on functional changes in the cysts and their relationship to breast cancer.

scholarly journals The serological grouping system forSerpulina (Treponema) hyodysenteriae

1992 ◽  
Vol 109 (2) ◽  
pp. 255-263 ◽  
Author(s):  
T. T. A. Lau ◽  
D. J. Hampson
Keyword(s):  
Silver Staining ◽  
Gel Electrophoresis ◽  
Lipid A ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cross Reactivity ◽  
Side Chain ◽  
Sodium Dodecylsulphate ◽  
Western Immunoblotting ◽  
The Core ◽  
Treponema Hyodysenteriae

SUMMARYLipopolysaceharide from serostrains ofSerpulina (Treponema) hyodysenteriaefor serogroups A to I was characterized using sodium dodecylsulphate polyacrylamide gel electrophoresis and silver staining. All strains had lipo-polysaccharide components ranging from 10 to 16 kDa that represented lipid A-core polysaccharide regions, and short O-antigen side chain were also recognized in certain immunoblots.Serological reactions between lipopolysaceharide and antisera against each of these serostrains were examined by Western immunoblotting. There was relatively little antigenic cross-reactivity between LPS from the nine strains, thus confirming their suitability as serostrains.Using cross-absorbed sera, isolates within serogroups A and E were shown to possess unique epitopes on the core lipopolysaceharide, distinct from serogroup reactivities. These isolates were therefore identified as serovars within the serogroups.This study confirmed the usefulness of the serotyping scheme forS. hyodysenteriae, in which the bacteria can be placed into serogroups using unabsorbed sera, and into serovars within these using cross-absorbed sera.

Lipopolysaccharide and proteins of the cell envelope of Vibrio marinus, a marine bacterium

1973 ◽  
Vol 19 (10) ◽  
pp. 1211-1217 ◽  
Author(s):  
Carl F. Deneke ◽  
R. R. Colwell
Keyword(s):  
Fatty Acids ◽  
Gel Electrophoresis ◽  
Marine Bacterium ◽  
Lipid A ◽  
Cell Envelope ◽  
Polyacrylamide Gel Electrophoresis ◽  
Amino Sugar ◽  
High Ratio ◽  
Side Chain ◽  
Total Fatty Acids

Lipopolysaccharides isolated from the marine bacterium Vibrio marinus strain PS-207 were found to be similar to the lipopolysaccharides of R mutants of enteric organisms, with respect to extraction characteristics, percentage of lipid A (61%), and sugars of the polysaccharide side chain (glucose and heptose). A high ratio (2:1) of phosphate to amino sugar was found in the lipid A. Hydroxy fatty acids constituted only 14% of the total fatty acids of the lipid A fraction, whereas branched and straight-chain fatty acids were present in greater abundance. The major envelope proteins of V. marinus strain PS-207 fell into three molecular weight classes determined by SDS gel electrophoresis. Numerous protein species were observed in urea – acetic polyacrylamide gel electrophoresis preparations.

THE STRUCTURES OF THE CORE-LIPID A REGIONS OF LIPOPOLYSACCHARIDES FROM SERRATIA MARCESCENS

2002 ◽  
Author(s):  
Evgeny Vinogradov ◽  
Buko Lindner ◽  
Guntram Seltmann ◽  
Joanna Radziejewska-Lebrecht ◽  
Otto Holst
Keyword(s):  
Serratia Marcescens ◽  
Lipid A ◽  
The Core ◽  
Core Lipid

scholarly journals Human erythrocyte antigens: II. The In(Lu) gene regulates expression of an antigen on an 80-kilodalton protein of human erythrocytes

Blood ◽  
1984 ◽  
Vol 64 (3) ◽  
pp. 599-606 ◽  
Author(s):  
MJ Telen ◽  
TJ Palker ◽  
BF Haynes
Keyword(s):  
Monoclonal Antibody ◽  
Western Blot ◽  
Erythrocyte Membrane ◽  
Human Erythrocyte ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein A ◽  
Sodium Dodecyl ◽  
Two Dimensions ◽  
Antigen Activity

Abstract We have previously shown that a murine monoclonal antibody (A3D8) identifies a human erythrocyte protein antigen whose expression is regulated by the Lutheran inhibitor [In(Lu)] gene. In the present study, we demonstrated by immunoprecipitation and Western blot techniques that the antigen defined by A3D8 was on an 80-kD erythrocyte membrane protein. A second 170-kD protein was coprecipitated with the 80-kD protein but failed to show antigen activity by Western blot analysis. The 170-kD protein, when analyzed by sodium dodecyl sulfate- polyacrylamide gel electrophoresis in two dimensions, was composed of 50- and 30-kD disulfide-linked subunits. In(Lu) Lu[a-b-) erythrocytes differed from Lu(a+b+) or Lu(a-b+) erythrocytes in that In(Lu) deoxycholate erythrocyte membrane extracts contained trace amounts of immunoprecipitable 80-kD protein compared with detergent-solubilized erythrocyte membrane extracts prepared from Lu(a+b+) or Lu(a-b+) subjects.

Structure of the O-chain of the lipopolysaccharide of Haemophilus pleuropneumoniae serotype 1

1986 ◽  
Vol 64 (12) ◽  
pp. 1317-1325 ◽  
Author(s):  
Eleonora Altman ◽  
Jean-Robert Brisson ◽  
Malcolm B. Perry
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Lipid A ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Magnetic Resonance Studies ◽  
Serotype 1 ◽  
Structure Formula ◽  
Phenol Water

By phenol-water extraction an aqueous-phase soluble cellular lipopolysaccharide was isolated from Haemophilus pleuropneumoniae serotype 1. It was shown by sodium dodecyl sulfate – polyacrylamide gel electrophoresis, hydrolysis, methylation, and both one- and two-dimensional 1H and 13C nuclear magnetic resonance studies to be an S-type lipopolysaccharide, which could be cleaved to yield a lipid A and an O-chain polysaccharide identified as a high molecular weight branched polymer of a tetrasaccharide repeating unit having the structure:[Formula: see text]

scholarly journals Identification of a 64 kDa heparan sulphate proteoglycan core protein from human lung fibroblast plasma membranes with a monoclonal antibody

1987 ◽  
Vol 247 (3) ◽  
pp. 765-771 ◽  
Author(s):  
H de Boeck ◽  
V Lories ◽  
G David ◽  
J J Cassiman ◽  
H van den Berghe
Keyword(s):  
Molecular Mass ◽  
Gel Electrophoresis ◽  
Human Lung ◽  
Core Protein ◽  
Heparan Sulphate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Apparent Molecular Mass ◽  
Lung Fibroblasts ◽  
Human Lung Fibroblasts ◽  
The Core

Human lung fibroblasts produce heparan sulphate proteoglycans (HSPG) that are associated with the plasma membrane. A monoclonal-antibody (Mab)-secreting hybridoma, S1, was produced by fusion of SP 2/0-AG 14 mouse myeloma cells with spleen cells from mice immunized with partially purified cellular HSPG fractions. The HSPG character of the material carrying the epitope recognized by Mab S1 was demonstrated by: (i) the co-purification of the S1 epitope with the membrane HSPG of human lung fibroblasts; (ii) the decrease in size of the material carrying the S1 epitope upon treatment with heparinase or heparitinase, and the resistance of this material to heparinase treatment after N-desulphation. The S1 epitope appears to be part of the core protein, since it was destroyed by proteinase treatment and by disulphide-bond reduction, but not by treatments that depolymerize the glycosaminoglycan chains and N-linked oligosaccharide chains. Polyacrylamide-gel electrophoresis of non-reduced heparitinase-digested membrane HSPG followed by Western blotting and immunostaining with Mab S1 revealed a single band with apparent molecular mass of 64 kDa. Membrane proteoglycans isolated from detergent extracts or from 4 M-guanidinium chloride extracts of the cells yielded similar results. Additional digestion with N-glycanase lowered the apparent molecular mass of the immunoreactive material to 56 kDa, suggesting that the core protein also carries N-linked oligosaccharides. Fractionation of 125I-labelled membrane HSPG by immuno-affinity chromatography on immobilized Mab S1, followed by heparitinase digestion and polyacrylamide-gel electrophoresis of the bound material, yielded a single labelled band with apparent molecular mass 64 kDa. Treatment with dithiothreitol caused a slight increase in apparent molecular mass, suggesting that the core protein of this membrane proteoglycan of a single subunit containing (an) intrachain disulphide bond(s).

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