Bile and Unsaturated Fatty Acids Inhibit the Binding of Cholera Toxin and Escherichia coli Heat-Labile Enterotoxin to GM1 Receptor

ABSTRACT Cholera toxin (CT) is an archetypal bacterial toxin that binds with a high affinity to the receptor ganglioside GM1 on the intestinal epithelial surface and that causes the severe watery diarrhea characteristic of the disease cholera. Blockage of the interaction of CT with the GM1 receptor is an attractive approach for therapeutic intervention. We report here that crude bile prevents the interaction of CT with GM1 and reduces CT-mediated fluid accumulation in the rabbit intestine. The unsaturated fatty acids detected in crude bile, arachidonic, linoleic, and oleic acids, were found to be the most effective. Crude bile and the unsaturated fatty acids interacted with CT but not GM1 to prevent CT-GM1 binding. Neither crude bile nor the unsaturated fatty acids had any effect on the subunit structure of CT. The binding of CT to unsaturated fatty acids resulted in a shift of the apparent pI of CT from 6.8 to 8.2 and a marked decrease in intrinsic fluorescence. The Kd was calculated from fluorescence quenching assays. It was demonstrated by the rabbit ileal loop model that practically no fluid accumulated in the intestinal loops when CT was administered together with inhibitory concentrations of linoleic acid. The bile present in the intestine was sufficient to inhibit the activity of up to 300 ng CT. Bile and unsaturated fatty acids also inhibited the binding of Escherichia coli heat-labile enterotoxin (LT) to GM1, and no fluid accumulation was observed in rabbit ileal loops when LT was administered together with linoleic acid.

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A New Member of the Escherichia coli fad Regulon: Transcriptional Regulation of fadM (ybaW)

Journal of Bacteriology ◽

10.1128/jb.00835-09 ◽

2009 ◽

Vol 191 (20) ◽

pp. 6320-6328 ◽

Cited By ~ 36

Author(s):

Youjun Feng ◽

John E. Cronan

Keyword(s):

Escherichia Coli ◽

Fatty Acids ◽

Linoleic Acid ◽

Conjugated Linoleic Acid ◽

Unsaturated Fatty Acids ◽

Negative Regulator ◽

Receptor Protein ◽

Conjugated Linoleic Acid Isomer ◽

Linoleic Acid Isomer

ABSTRACT Recently, Nie and coworkers (L. Nie, Y. Ren, A. Janakiraman, S. Smith, and H. Schulz, Biochemistry 47:9618-9626, 2008) reported a new Escherichia coli thioesterase encoded by the ybaW gene that cleaves the thioester bonds of inhibitory acyl-coenzyme A (CoA) by-products generated during β-oxidation of certain unsaturated fatty acids. These authors suggested that ybaW expression might be regulated by FadR, the repressor of the fad (fatty acid degradation) regulon. We report mapping of the ybaW promoter and show that ybaW transcription responded to FadR in vivo. Moreover, purified FadR bound to a DNA sequence similar to the canonical FadR binding site located upstream of the ybaW coding sequence and was released from the promoter upon the addition of long-chain acyl-CoA thioesters. We therefore propose the designation fadM in place of ybaW. Although FadR regulation of fadM expression had the pattern typical of fad regulon genes, its modulation by the cyclic AMP (cAMP) receptor protein-cAMP complex (CRP-cAMP) global regulator was the opposite of that normally observed. CRP-cAMP generally acts as an activator of fad gene expression, consistent with the low status of fatty acids as carbon sources. However, glucose growth stimulated fadM expression relative to acetate growth, as did inactivation of CRP-cAMP, indicating that the complex acts as a negative regulator of this gene. The stimulation of fadM expression seen upon deletion of the gene encoding adenylate cyclase (Δcya) was reversed by supplementation of the growth medium with cAMP. Nie and coworkers also reported that growth on a conjugated linoleic acid isomer yields much higher levels of FadM thioesterase activity than does growth on oleic acid. In contrast, we found that the conjugated linoleic acid isomer was only a weak inducer of fadM expression. Although the gene is not essential for growth, the high basal level of fadM expression under diverse growth conditions suggests that the encoded thioesterase has functions in addition to β-oxidation.

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Explore the gene network regulating the composition of fatty acids in cottonseed

BMC Plant Biology ◽

10.1186/s12870-021-02952-4 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Lihong Ma ◽

Xinqi Cheng ◽

Chuan Wang ◽

Xinyu Zhang ◽

Fei Xue ◽

...

Keyword(s):

Fatty Acids ◽

Fatty Acid ◽

Linoleic Acid ◽

Linolenic Acid ◽

Gene Network ◽

Unsaturated Fatty Acids ◽

Saturated Fatty Acids ◽

Accumulation Pattern ◽

Time Points ◽

Expression Characteristics

Abstract Background Cottonseed is one of the major sources of vegetable oil. Analysis of the dynamic changes of fatty acid components and the genes regulating the composition of fatty acids of cottonseed oil is of great significance for understanding the biological processes underlying biosynthesis of fatty acids and for genetic improving the oil nutritional qualities. Results In this study, we investigated the dynamic relationship of 13 fatty acid components at 12 developmental time points of cottonseed (Gossypium hirsutum L.) and generated cottonseed transcriptome of the 12 time points. At 5–15 day post anthesis (DPA), the contents of polyunsaturated linolenic acid (C18:3n-3) and saturated stearic acid (C18:0) were higher, while linoleic acid (C18:2n-6) was mainly synthesized after 15 DPA. Using 5 DPA as a reference, 15,647 non-redundant differentially expressed genes were identified in 10–60 DPA cottonseed. Co-expression gene network analysis identified six modules containing 3275 genes significantly associated with middle-late seed developmental stages and enriched with genes related to the linoleic acid metabolic pathway and α-linolenic acid metabolism. Genes (Gh_D03G0588 and Gh_A02G1788) encoding stearoyl-ACP desaturase were identified as hub genes and significantly up-regulated at 25 DPA. They seemed to play a decisive role in determining the ratio of saturated fatty acids to unsaturated fatty acids. FAD2 genes (Gh_A13G1850 and Gh_D13G2238) were highly expressed at 25–50 DPA, eventually leading to the high content of C18:2n-6 in cottonseed. The content of C18:3n-3 was significantly decreased from 5 DPA (7.44%) to 25 DPA (0.11%) and correlated with the expression characteristics of Gh_A09G0848 and Gh_D09G0870. Conclusions These results contribute to our understanding on the relationship between the accumulation pattern of fatty acid components and the expression characteristics of key genes involved in fatty acid biosynthesis during the entire period of cottonseed development.

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The aerobic oxidation of unsaturated fatty acids and their esters: cobalt stearate-catalyzed oxidation of linoleic acid

The Journal of Lipid Research ◽

10.1016/s0022-2275(20)39024-6 ◽

1961 ◽

Vol 2 (2) ◽

pp. 152-160

Author(s):

F.W. Heaton ◽

N. Uri

Keyword(s):

Fatty Acids ◽

Linoleic Acid ◽

Unsaturated Fatty Acids ◽

Aerobic Oxidation ◽

Catalyzed Oxidation

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A genetically detoxified derivative of heat-labile Escherichia coli enterotoxin induces neutralizing antibodies against the A subunit.

Journal of Experimental Medicine ◽

10.1084/jem.180.6.2147 ◽

1994 ◽

Vol 180 (6) ◽

pp. 2147-2153 ◽

Cited By ~ 69

Author(s):

M Pizza ◽

M R Fontana ◽

M M Giuliani ◽

M Domenighini ◽

C Magagnoli ◽

...

Keyword(s):

Escherichia Coli ◽

Cholera Toxin ◽

Neutralizing Antibodies ◽

Site Directed Mutagenesis ◽

Directed Mutagenesis ◽

E Coli ◽

Heat Labile ◽

A Subunit ◽

Adp Ribosyltransferase ◽

Derivatives Of

Escherichia coli enterotoxin (LT) and the homologous cholera toxin (CT) are A-B toxins that cause travelers' diarrhea and cholera, respectively. So far, experimental live and killed vaccines against these diseases have been developed using only the nontoxic B portion of these toxins. The enzymatically active A subunit has not been used because it is responsible for the toxicity and it is reported to induce a negligible titer of toxin neutralizing antibodies. We used site-directed mutagenesis to inactivate the ADP-ribosyltransferase activity of the A subunit and obtained nontoxic derivatives of LT that elicited a good titer of neutralizing antibodies recognizing the A subunit. These LT mutants and equivalent mutants of CT may be used to improve live and killed vaccines against cholera and enterotoxinogenic E. coli.

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Biochemical and structural characterization of the novel sialic acid-binding site of Escherichia coli heat-labile enterotoxin LT-IIb

Biochemical Journal ◽

10.1042/bcj20160575 ◽

2016 ◽

Vol 473 (21) ◽

pp. 3923-3936 ◽

Cited By ~ 3

Author(s):

Dani Zalem ◽

João P. Ribeiro ◽

Annabelle Varrot ◽

Michael Lebens ◽

Anne Imberty ◽

...

Keyword(s):

Escherichia Coli ◽

Binding Site ◽

Cholera Toxin ◽

Carbohydrate Binding ◽

Type I ◽

Type Ii ◽

E Coli ◽

B Subunit ◽

Heat Labile ◽

B Subunits

The structurally related AB5-type heat-labile enterotoxins of Escherichia coli and Vibrio cholerae are classified into two major types. The type I group includes cholera toxin (CT) and E. coli LT-I, whereas the type II subfamily comprises LT-IIa, LT-IIb and LT-IIc. The carbohydrate-binding specificities of LT-IIa, LT-IIb and LT-IIc are distinctive from those of cholera toxin and E. coli LT-I. Whereas CT and LT-I bind primarily to the GM1 ganglioside, LT-IIa binds to gangliosides GD1a, GD1b and GM1, LT-IIb binds to the GD1a and GT1b gangliosides, and LT-IIc binds to GM1, GM2, GM3 and GD1a. These previous studies of the binding properties of type II B-subunits have been focused on ganglio core chain gangliosides. To further define the carbohydrate binding specificity of LT-IIb B-subunits, we have investigated its binding to a collection of gangliosides and non-acid glycosphingolipids with different core chains. A high-affinity binding of LT-IIb B-subunits to gangliosides with a neolacto core chain, such as Neu5Gcα3- and Neu5Acα3-neolactohexaosylceramide, and Neu5Gcα3- and Neu5Acα3-neolactooctaosylceramide was detected. An LT-IIb-binding ganglioside was isolated from human small intestine and characterized as Neu5Acα3-neolactohexaosylceramide. The crystal structure of the B-subunit of LT-IIb with the pentasaccharide moiety of Neu5Acα3-neolactotetraosylceramide (Neu5Ac-nLT: Neu5Acα3Galβ4GlcNAcβ3Galβ4Glc) was determined providing the first information for a sialic-binding site in this subfamily, with clear differences from that of CT and LT-I.

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Fatty acid positional distribution (sn-2 fatty acids) and phospholipid composition in Chinese breast milk from colostrum to mature stage

British Journal Of Nutrition ◽

10.1017/s0007114518002994 ◽

2018 ◽

Vol 121 (1) ◽

pp. 65-73 ◽

Cited By ~ 9

Author(s):

Ke Wu ◽

Runying Gao ◽

Fang Tian ◽

Yingyi Mao ◽

Bei Wang ◽

...

Keyword(s):

Fatty Acids ◽

Fatty Acid ◽

Linoleic Acid ◽

Fatty Acid Profile ◽

Unsaturated Fatty Acids ◽

Fatty Acid Content ◽

Positional Distribution ◽

Phospholipid Composition ◽

Mature Stage ◽

Feeding Time

AbstractThis study quantified the fatty acid profile with emphasis on the stereo-specifically numbered (sn) 2 positional distribution in TAG and the composition of main phospholipids at different lactation stages. Colostrum milk (n 70), transitional milk (n 96) and mature milk (n 82) were obtained longitudinally from healthy lactating women in Shanghai. During lactation, total fatty acid content increased, with SFA dominating in fatty acid profile. A high ratio of n-6:n-3 PUFA was observed as 11:1 over lactation due to the abundance of linoleic acid in Chinese human milk. As the main SFA, palmitic acid showed absolute sn-2 selectivity, while oleic acid, linoleic acid and α-linolenic acid, the main unsaturated fatty acids, were primarily esterified at the sn-1 and sn-3 positions. Nervonic acid and C22 PUFA including DHA were more enriched in colostrum with an sn-2 positional preference. A total of three dominant phospholipids (phosphatidylethanolamine (PE), phosphatidylcholine (PC) and sphingomyelin (SM)) were analysed in the collected samples, and each showed a decline in amount over lactation. PC was the dominant compound followed by SM and PE. With prolonged breast-feeding time, percentage of PE in total phospholipids remained constant, but PC decreased, and SM increased. Results from this study indicated a lipid profile different from Western reports and may aid the development of future infant formula more suitable for Chinese babies.

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Escherichia coli Heat-Labile Enterotoxin B Subunit Is a More Potent Mucosal Adjuvant than Its Closely Related Homologue, the B Subunit of Cholera Toxin

Infection and Immunity ◽

10.1128/iai.69.5.3476-3482.2001 ◽

2001 ◽

Vol 69 (5) ◽

pp. 3476-3482 ◽

Cited By ~ 57

Author(s):

Douglas G. Millar ◽

Timothy R. Hirst ◽

Denis P. Snider

Keyword(s):

Escherichia Coli ◽

Cholera Toxin ◽

Lymphocyte Proliferation ◽

Vaccine Adjuvants ◽

B Subunit ◽

Heat Labile ◽

Antibody Titers ◽

Enterotoxin B ◽

Draining Lymph Nodes ◽

Stimulation Of

ABSTRACT Although cholera toxin (Ctx) and Escherichia coliheat-labile enterotoxin (Etx) are known to be potent mucosal adjuvants, it remains controversial whether the adjuvanticity of the holotoxins extends to their nontoxic, receptor-binding B subunits. Here, we have systematically evaluated the comparative adjuvant properties of highly purified recombinant EtxB and CtxB. EtxB was found to be a more potent adjuvant than CtxB, stimulating responses to hen egg lysozyme when the two were coadministered to mice intranasally, as assessed by enhanced serum and secretory antibody titers as well as by stimulation of lymphocyte proliferation in spleen and draining lymph nodes. These results indicate that, although structurally very similar, EtxB and CtxB have strikingly different immunostimulatory properties and should not be considered equivalent as prospective vaccine adjuvants.

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Inhibition of steroid 5α-reductase by specific aliphatic unsaturated fatty acids

Biochemical Journal ◽

10.1042/bj2850557 ◽

1992 ◽

Vol 285 (2) ◽

pp. 557-562 ◽

Cited By ~ 126

Author(s):

T Liang ◽

S Liao

Keyword(s):

Fatty Acids ◽

Oleic Acid ◽

Linoleic Acid ◽

Linolenic Acid ◽

Unsaturated Fatty Acids ◽

Binding Assay ◽

Reductase Activity ◽

Undecylenic Acid ◽

Gamma Linolenic Acid ◽

Enzymic Assay

Human or rat microsomal 5 alpha-reductase activity, as measured by enzymic conversion of testosterone into 5 alpha-dihydrotestosterone or by binding of a competitive inhibitor, [3H]17 beta-NN-diethulcarbamoyl-4-methyl-4-aza-5 alpha-androstan-3-one ([3H]4-MA) to the reductase, is inhibited by low concentrations (less than 10 microM) of certain polyunsaturated fatty acids. The relative inhibitory potencies of unsaturated fatty acids are, in decreasing order: gamma-linolenic acid greater than cis-4,7,10,13,16,19-docosahexaenoic acid = cis-6,9,12,15-octatetraenoic acid = arachidonic acid = alpha-linolenic acid greater than linoleic acid greater than palmitoleic acid greater than oleic acid greater than myristoleic acid. Other unsaturated fatty acids such as undecylenic acid, erucic acid and nervonic acid, are inactive. The methyl esters and alcohol analogues of these compounds, glycerols, phospholipids, saturated fatty acids, retinoids and carotenes were inactive even at 0.2 mM. The results of the binding assay and the enzymic assay correlated well except for elaidic acid and linolelaidic acid, the trans isomers of oleic acid and linoleic acid respectively, which were much less active than their cis isomers in the binding assay but were as potent in the enzymic assay. gamma-Linolenic acid had no effect on the activities of two other rat liver microsomal enzymes: NADH:menadione reductase and glucuronosyl transferase. gamma-Linolenic acid, the most potent inhibitor tested, decreased the Vmax. and increased Km values of substrates, NADPH and testosterone, and promoted dissociation of [3H]4-MA from the microsomal reductase. gamma-Linolenic acid, but not the corresponding saturated fatty acid (stearic acid), inhibited the 5 alpha-reductase activity, but not the 17 beta-dehydrogenase activity, of human prostate cancer cells in culture. These results suggest that unsaturated fatty acids may play an important role in regulating androgen action in target cells.

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Profile of ingested fatty acids and in the duodenal digest of steers fed different diets

Revista Brasileira de Zootecnia ◽

10.1590/s1516-35982010001100025 ◽

2010 ◽

Vol 39 (11) ◽

pp. 2502-2511 ◽

Cited By ~ 4

Author(s):

Luís Fernando Glasenapp de Menezes ◽

Gilberto Vilmar Kozloski ◽

João Restle ◽

Ivan Luiz Brondani ◽

Raul Dirceu Pazdiora ◽

...

Keyword(s):

Fatty Acids ◽

Linoleic Acid ◽

Conjugated Linoleic Acid ◽

Unsaturated Fatty Acids ◽

Saturated Fatty Acids ◽

Latin Square ◽

Tropical Pasture ◽

Ruminal Fermentation ◽

Omega 3 ◽

Conventional Diet

It was evaluated in this study the effect of the type of the diet on duodenal flow of long-chain fatty acids in steers. The tested diets were the following: conventional (feedlot diet composed of 60% corn silage and 40% of concentrate); winter forage silage - rye grass (Lolium multiflorum, Lam); or tropical forage silage - association of millet (Pennisetum americanum, Leeke + alexander grass, Brachiaria plantaginea). Six Charolais × Nellore crossbred steers with cannulas in duodenum were used in a 3 × 3 double Latin square. Dry material intake was similar among the groups (mean of 4,037 g/day), but the intake of total fatty acids and saturated fatty acids were higher in the group fed tropical pasture silage. On the other hand, the animals which received the conventional diet consumed higher quantity of unsaturated fatty acids. Tropical pasture silage provided higher consumption of vacenic acid (C18:1 t-11) and the winter forage silage offered higher consumption of conjugated linoleic acid. The intake of omega-6 fatty acids was higher in the group fed conventional diet and for omega-3, intake was higher in the group fed tropical pasture diet. The total fatty acid flow in the duodenum was not affected by the diets, but in all treatments it was higher than the consumed one. The animals fed diet with concentrate show the greatest changes on the profile of fatty acids during the ruminal fermentation. Conventional diets provide the highest intake of unsaturated fatty acids and the highest availability of vacenic acid in the small intestine, but they do not increase the supply of intestinal conjugated linoleic acid.

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