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.

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.

Studies on the lipopolysaccharide of a virulent and an avirulent strain of Vibrio vulnificus

1990 ◽  
Vol 68 (2) ◽  
pp. 547-551 ◽  
Author(s):  
K. Bahrani ◽  
James D. Oliver
Keyword(s):  
Fatty Acids ◽  
Marine Bacterium ◽  
Lipid A ◽  
Vibrio Vulnificus ◽  
Virulent Strain ◽  
Chemical Constituents ◽  
Avirulent Strain ◽  
Membrane Phospholipids ◽  
Total Fatty Acids ◽  
Pattern Characteristic

Vibrio vulnificus is a marine bacterium associated with both primary septicemias and wound infections in humans. The lipopolysaccharides of a virulent and an avirulent strain of Vibrio vulnificus were compared with respect to their chemical constituents and electrophoretic characteristics. 2-Keto-3-deoxyoctonic acid, a normal constituent of the lipopolysaccharide of typical Enterobacteriaceae, was not found in the lipopolysaccharide of either strain. Hexadecenoate (C16:1) was the predominant fatty acid of the lipid A moiety of the lipopolysaccharides and of the membrane phospholipids of both strains. Hydroxy fatty acids composed 44% of the total fatty acids of the lipid A of the avirulent and 40% of those in the virulent strain. In addition, odd-numbered fatty acids were detected in both lipopolysaccharides. The electrophoretic profile was similar for both strains, but demonstrated no "ladder-like" pattern characteristic of "smooth" lipopolysaccharides. The result of this study showed no significant differences between the lipopolysaccharides of the virulent and avirulent strains of Vibrio vulnificus. The possible role for lipopolysaccharide in pathogenesis of Vibrio vulnificus infections is discussed.Key words: Vibrio, lipopolysaccharide, pathogenesis.

Comparative Immunological and Chemical Analysis of Lipids and Carotenoids of the D1-Peptide and of the Light-Harvesting-Complex of Photosystem II of Nicotiana tabacum

1999 ◽  
Vol 54 (3-4) ◽  
pp. 199-208 ◽  
Author(s):  
Anette Gasser ◽  
Stefan Raddatz ◽  
Alfons Radunz ◽  
Georg H. Schmid
Keyword(s):  
Fatty Acids ◽  
Nicotiana Tabacum ◽  
Photosystem Ii ◽  
Western Blot ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Light Harvesting Complex ◽  
Total Fatty Acids ◽  
Monospecific Antisera ◽  
Β Carotene

The light-harvesting-complex (LHCP) was isolated from photosystem II of Nicotiana tabacum var. John William’s Broadleaf by means of the detergent acetyl-β-ᴅ-glucopyranoside and fractionating centrifugation. The D1-peptide of photosystem II was isolated as a dimer with the molecular mass of 66 kDa from the chlorophyll-deficient tobacco mutant N. tabacum Su/su by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis. Both preparations were characterized by means of the Western blot procedure using monospecific antisera to the proteins of photosystem II and monospecific antisera to lipids with which the lipids bound to peptides were determined. In parallel to this, lipids bound to the isolated LHCP-complex and to the isolated D1-peptide were determined by lipido-chemical methods. The extraction of the isolated core peptide D1 with a mixture of boiling methanol and chloroform and subsequent HPLC-chromatography showed that in the D1-peptide isolated via SDS-polyacrylamide gel electrophoresis, monogalactolipids, phosphatidylglycerol and sulfolipid molecules are bound in the molar ratio 1:3:17. By means of the immunological procedure of Western blotting we were able to show that the 66 kDa band of the isolated dimeric D1 reacts positively only with the antisera to monogalactolipid, sulfolipid, β-carotene and violaxanthin. With the antiserum to digalactolipid and that to phosphatidylglycerol a positive reaction is only observed if the preparation used in the Western blot is not the isolated D1-peptide but a ‘‘total” photosystem Il-preparation. The lipid extraction of the LHCP-complex and the subsequent analysis by thin-layer chromatography led to the result that the isolated LHCP-complex contained in bound form 3 molecules monogalactolipid, 1 molecule of digalactolipid, 1 molecule of phosphatidylglycerol and 1 molecule of lutein. Less than 1 molecule of sulfolipid, β-carotene, neoxanthin and violaxanthin are found. In the Western blot analysis only the antiserum to monogalactolipid and phosphatidylglycerol and among the carotenoid antisera only the antisera to β-carotene, violaxanthin and to neoxanthin reacted. With the antisera to the digalactolipid, to the sulfolipid and the antisera to the xanthophylls, namely to lutein and neoxanthin, a positive reaction occurred only if the material used in the Western Blot was the “total” photosystem Il-preparation. By gas chromatography of the fatty acids of the isolated peptide fractions it was shown that, compared to the lipids of photosystem II and of the thylakoid membrane, in lipids of the isolated D1-peptide and of the LHCP-complex the saturation degree of fatty acids is strongly increased. Whereas palmitic acid in chloroplast lipids makes up for only 11% of the fatty acids, this saturated fatty acid increases in the lipids of the LHCP to 20% and makes up for 74% of total fatty acids in the lipids of the D1-peptide. Linoleic and linolenic acids are completely absent and oleic acid makes up for 14% of total fatty acids. In contrast to the lipids of the thylakoid membrane, the lipids bound to proteins/peptides are characterized by a strongly saturated character.

Studies of the cell envelope of Vibrio parahaemolyticus

1973 ◽  
Vol 19 (2) ◽  
pp. 241-245 ◽  
Author(s):  
Carl F. Deneke ◽  
R. R. Colwell
Keyword(s):  
Fatty Acids ◽  
Vibrio Parahaemolyticus ◽  
Lipid A ◽  
Cell Envelope ◽  
Extraction Procedure ◽  
Amino Sugar ◽  
Molar Ratio ◽  
Gas Liquid Chromatography ◽  
Phenol Extraction

Components of the cell envelope of Vibrio parahaemolyticus were investigated. Vibrio parahaemolyticus is an estuarine microorganism associated with diseases of marine and estuarine animals and seafood-borne enteritis in man. Purified lipopolysaccharide (LPS), isolated using a 45% phenol extraction procedure, was found to contain lipid A fraction to 27% of the LPS by weight. In the lipid A fraction, glucosamine was the only amino sugar to be present and a high molar ratio of phosphate to amino sugar (2.5:1) was noted. Two hydroxy fatty acids, hydroxydodecanoic and hydroxymyristic, were identified among the fatty acids by gas–liquid chromatography. A role of the lipopolysaccharides in the salt requirement of marine bacteria is suggested.

scholarly journals The acylation of megakaryocyte proteins: glycoprotein IX is primarily myristoylated while glycoprotein Ib is palmitoylated

Blood ◽  
1996 ◽  
Vol 87 (4) ◽  
pp. 1377-1384 ◽  
Author(s):  
PK Schick ◽  
J Walker
Keyword(s):  
Fatty Acids ◽  
Myristic Acid ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Amide Bond ◽  
Endogenous Protein ◽  
Sds Page ◽  
Palmitic Acids ◽  
Protein Acylation

The acylation of megakaryocyte proteins was studied with special emphasis on the myristoylation and palmitoylation of the glycoprotein (GP) Ib complex. Guinea pig megakaryocytes were purified and separated into subpopulations at different phases of maturation. Cells were incubated with [3H]myristate, [3H]palmitate, or [3H]acetate to study endogenous protein acylation. Cycloheximide was used to distinguish between cotranslational and posttranslational acylation and hydroxylamine to distinguish between thioester and amide linkages. After incubations, delipidated proteins or GPIb complex subunits, immunoprecipitated with PG-1, AN-51 or FMC-25 monoclonal antibody, were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and assessed by fluorography. Radiolabeled fatty acids bound to GPIX and GPIb were also analyzed by high pressure liquid chromatography (HPLC) and scintillation spectrometry. With [3H]myristic acid and [3H]acetate, GPIX was found to be a major myristoylated protein in megakaryocytes and CHRF-288 cells. Myristic acid was linked to GPIX by an amide bond, and this process occurred cotranslationally. With [3H]acetate, GPIb was primarily palmitoylated, but with [3H]myristate, GPIb was acylated with about equal mounts of myristic acid and palmitic acids. Both fatty acids were linked to GPIb by thioester bonds, and acylation was posttranslational. The myristoylation of GPIX while the palmitoylation of GPIb occurred throughout megakaryocyte maturation. Myristoylation and palmitoylation may have different functions relevant to the assembly of the GPIb complex in megakaryocytes.

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]

Studies on the chemical constitution of cell-wall lipopolysaccharides from Neisseria perflava

1968 ◽  
Vol 46 (10) ◽  
pp. 1175-1184 ◽  
Author(s):  
G. A. Adams ◽  
M. Kates ◽  
D. H. Shaw ◽  
M. Yaguchi
Keyword(s):  
Fatty Acids ◽  
Lipid A ◽  
Glycosidic Bonds ◽  
Chemical Constitution ◽  
Total Fatty Acids ◽  
Neutral Sugars ◽  
A Chain ◽  
Ethanolamine Phosphate ◽  
Neisseria Perflava ◽  
Palmitic Acids

Lipopolysaccharides (LPS) were prepared from Neisseria perflava cells and separated into chloroform-soluble and chloroform-insoluble fractions in about equal proportions. The preparations were almost free from protein and contained only traces of peptide material. Both LPS fractions contained 3-deoxyoctulosonic acid (KDO), lipid A, glucose, rhamnose, heptose, glucosamine, galactosamine, ethanolamine, phosphate, and fatty acids. Both fractions had the same proportions of hexosamines and ethanolamine and the same fatty acid components (chiefly β-hydroxymyristic, β-faydroxylauric, octadecenoic, and palmitic acids), but differed markedly in their relative molar proportions of KDO to neutral sugars and of neutral sugars to total fatty acids. Equimolar proportions of hexosamine, ethanolamine, and phosphate in the lipid A moieties suggested that ethanolamine phosphate was linked to glucosamine, probably at the C-6 position. Fatty acids were bound in the LPS by both amide and ester linkages. Heptose units were linked linearly by 1 → 3 glycosidic bonds. About 25% of the glucose units were linked 1 → 3 in a chain; other glucose units were branched as were the rhamnose units. The proportions and arrangement of the heptose, KDO, and lipid A components suggest that the backbone part of the N. perflava lipopolysaccharide is very similar to that found in the Salmonella-Escherichia group.

Lipopolysaccharides of Legionella erythra and Legionella oakridgensis

1994 ◽  
Vol 40 (8) ◽  
pp. 666-671 ◽  
Author(s):  
Anders Sonesson ◽  
Erik Jantzen ◽  
Torill Tangen ◽  
Ulrich Zähringer
Keyword(s):  
Fatty Acids ◽  
Lipid A ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Long Chain Fatty Acids ◽  
Glycerol Phosphate ◽  
Dioic Acid ◽  
Glucose Phosphate ◽  
Type Character ◽  
Long Chain

The chemical composition of lipopolysaccharides from Legionella erythra and Legionella oakridgensis was analysed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed both lipopolysaccharides to have a smooth-type character. The polysaccharide part of both lipopolysaccharides contained D-mannose, D-glucose, D-glycero-D-mannoheptose, L-glycero-D-manno-heptose, 2-keto-3-deoxyoctonic acid, L-fucosamine, D-glucosamine, and glucosamine phosphate. In addition, L-rhamnose, glycerol phosphate, and glucose phosphate were identified in the polysaccharide part of L. erythra lipopolysaccharide. The main sugar identified in the lipid A part of both lipopolysaccharides, 2,3-diamino-2,3-dideoxy-D-glucose, was found to be substituted with a complex fatty acid composition including at least 16 different amide-linked 3-hydroxy fatty acids. Both lipopolysaccharides contained nonhydroxy fatty acids and the uncommon 27-oxo-octacosanoic acid, 29-oxotriacontanoic acid, and 27-hydroxyoctacosanoic acid. The lipopolysaccharide of L. oakridgensis also contained 29-hydroxytriacontanoic acid. The dioic long-chain acids heptacosane-1,27-dioic acid and nonacosane-1,29-dioic acid were only present in the lipopolysaccharide of L. erythra.Key words: taxonomy, long-chain fatty acids, chemical analysis, 2,3-diamino-2,3-dideoxy-D-glucose.

Comparison of the cell envelope proteins of Micrococcus cryophilus with those of Neisseria and Branhamella species

1976 ◽  
Vol 22 (2) ◽  
pp. 309-312 ◽  
Author(s):  
R. R. B. Russell ◽  
I. J. McDonald
Keyword(s):  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Gel Electrophoresis ◽  
Cell Envelope ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Composition ◽  
Polyacrylamide Gel ◽  
Outer Membranes ◽  
Branhamella Catarrhalis

In an attempt to elucidate the relation between Micrococcus cryophilus, Neisseria caviae, Neisseria ovis, and Branhamella catarrhalis, fractions derived from outer membranes of a strain of each organism were examined for protein composition by SDS – polyacrylamide gel electrophoresis. Micrococcus cryophilus outer membrane protein showed extensive similarities to that of N. ovis and contained a heat-modifiable protein which behaved almost identically with the corresponding bands previously shown to exist in N. caviae and N. oris. Branhamella catarrhalis protein was distinctly different from those of M. cryophilus and the two 'false neisserias' N. caviae and N. oris.

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