Lipopolysaccharide

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Structure of a lipopolysaccharide
Lipopolysaccharides (LPS), also known as lipoglycans, are large molecules consisting of a
lipid and a polysaccharide joined by a covalent bond; they are found in the outer membrane
of Gram-negative bacteria, act as endotoxins and elicit strong immune responses in animals.
Contents
[hide]
1 Functions
2 Composition
o 2.1 O-antigen
o 2.2 Core
o 2.3 Lipid A
3 Biosynthesis and Transport
4 LPS modifications
5 Variability and effect upon specificity
6 Immune response
7 See also
8 References
9 External links
[edit] Functions
LPS is the major component of the outer membrane of Gram-negative bacteria, contributing
greatly to the structural integrity of the bacteria, and protecting the membrane from certain
kinds of chemical attack. LPS also increases the negative charge of the cell membrane and
helps stabilize the overall membrane structure. It is of crucial importance to gram-negative
bacteria, whose death results if it is mutated or removed. LPS is an endotoxin, and induces a
strong response from normal animal immune systems. It has also been implicated in non-
pathogenic aspects of bacterial ecology, including surface adhesion, bacteriophage
sensitivity, and interactions with predators such as amoebae.
LPS is required for the proper conformation of Omptin activity; however, smooth LPS will
sterically hinder omptins.
LPS acts as the prototypical endotoxin because it binds the CD14/TLR4/MD2 receptor
complex, which promotes the secretion of pro-inflammatory cytokines in many cell types, but
especially in macrophages and B cells. In Immunology, the term "LPS challenge" refers to
the process of exposing a subject to an LPS that may act as a toxin.
LPS is also an exogenous pyrogen (external fever-inducing substance).
Being of crucial importance to gram-negative bacteria, these molecules make candidate
targets for new antimicrobial agents.
Some researchers doubt reports of generalized toxic effects attributed to all
lipopolysaccharides, in particular, for cyanobacteria.
[1]

[edit] Composition


The saccharolipid Kdo
2
-Lipid A. Glucosamine residues in blue, Kdo residues in red, acyl
chains in black and phosphate groups in green.
It comprises three parts:
1. O antigen (or O polysaccharide)
2. Core oligosaccharide
3. Lipid A
[edit] O-antigen
A repetitive glycan polymer contained within an LPS is referred to as the O antigen, O
polysaccharide, or O side-chain of the bacteria. The O antigen is attached to the core
oligosaccharide, and comprises the outermost domain of the LPS molecule. The composition
of the O chain varies from strain to strain. For example, there are over 160 different O
antigen structures produced by different E. coli strains.
[2]
The presence or absence of O
chains determines whether the LPS is considered rough or smooth. Full-length O-chains
would render the LPS smooth, whereas the absence or reduction of O-chains would make the
LPS rough.
[3]
Bacteria with rough LPS usually have more penetrable cell membranes to
hydrophobic antibiotics, since a rough LPS is more hydrophobic.
[4]
O antigen is exposed on
the very outer surface of the bacterial cell, and, as a consequence, is a target for recognition
by host antibodies.
[edit] Core
Main article: Core oligosaccharide
The Core domain always contains an oligosaccharide component that attaches directly to
lipid A and commonly contains sugars such as heptose and 3-deoxy-D-mannooctulosonic
Acid (also known as KDO, keto-deoxyoctulosonate).
[5]
The LPS Cores of many bacteria also
contain non-carbohydrate components, such as phosphate, amino acids, and ethanolamine
substitutents.
[edit] Lipid A
Main article: Lipid A
Lipid A is, in normal circumstances, a phosphorylated glucosamine disaccharide decorated
with multiple fatty acids. These hydrophobic fatty acid chains anchor the LPS into the
bacterial membrane, and the rest of the LPS projects from the cell surface. The lipid A
domain is responsible for much of the toxicity of Gram-negative bacteria. When bacterial
cells are lysed by the immune system, fragments of membrane containing lipid A are released
into the circulation, causing fever, diarrhea, and possible fatal endotoxic shock (also called
septic shock).
[edit] Biosynthesis and Transport


LPS Final Assembly: O-antigen subunits are translocated across the inner membrane (by
Wzx) where they are polymerized (by Wzy, chain length determined by Wzz) and ligated (by
WaaL) on to complete Core-Lipid A molecules (which were translocated by MsbA).
[6]



LPS Transport: Completed LPS molecules are transported across the periplasm and outer
membrane by the proteins LptA, B, C, D, E, F, and G
[7]

[edit] LPS modifications
The making of LPS can be modified in order to present a specific sugar structure. Those can
be recognised by either other LPS (which enables to inhibit LPS toxins) or
glycosyltransferases that use those sugar structure to add more specific sugars. It has recently
been shown that a specific enzyme in the intestine (alkaline phosphatase) can detoxify LPS
by removing the two phosphate groups found on LPS carbohydrates.
[8]
This may function as
an adaptive mechanism to help the host manage potentially toxic effects of gram-negative
bacteria normally found in the small intestine.
[edit] Variability and effect upon specificity


Toll-like receptors of the innate immune system recognize LPS and trigger an immune
response.
O-antigens (the outer carbohydrates) are the most variable portion of the LPS molecule,
imparting the antigenic specificity. In contrast, lipid A is the most conserved part. However,
lipid A composition also may vary (e.g., in number and nature of acyl chains even within or
between genera). Some of these variations may impart antagonistic properties to these LPS.
For example Rhodobacter sphaeroides diphosphoryl lipid A (RsDPLA) is a potent antagonist
of LPS in human cells, but is an agonist in hamster and equine cells.
It has been speculated that conical Lipid A (e.g., from E. coli) are more agonistic, less conical
lipid A like those of Porphyromonas gingivalis may activate a different signal (TLR2 instead
of TLR4), and completely cylindrical lipid A like that of Rhodobacter sphaeroides is
antagonistic to TLRs.
[9][10]

Lipopolysaccharide gene clusters are highly variable between different strains, subspecies,
species of bacterial pathogens of plants and animals.
[11][12]

[edit] Immune response
LPS function has been under experimental research for several years due to its role in
activating many transcription factors. LPS challenge also produces many types of mediators
involved in septic shock. Humans are much more sensitive to LPS than other animals (e.g.,
mice). A dose of 1 g/kg induces shock in humans, but mice will tolerate a dose up to a
thousand times higher.
[13]
This may relate to differences in the level of circulating natural
antibodies between the two species.
[14][15]
Said et al. showed that LPS causes an IL-10-
dependent inhibition of CD4 T-cell expansion and function by up-regulating PD-1 levels on
monocytes which leads to IL-10 production by monocytes after binding of PD-1 by PD-L.
[16]

Bruce Beutler was awarded a portion of the 2011 Nobel Prize in Physiology or Medicine for
his work demonstrating that TLR4 is the LPS receptor.
[17][18]

[edit] See also
Endotoxin
Mucopolysaccharide
Nesfatin-1
[edit] References
1. ^ Stewart I, Schluter PJ, Shaw GR (2006). "Cyanobacterial lipopolysaccharides and human health a
review". Environ Health 5: 7. DOI:10.1186/1476-069X-5-7. PMC 1489932. PMID 16563160.
//www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=1489932.
2. ^ Christian Raetz and Chris Whitfield (2002) Lipopolysaccharide Endotoxins Annu. Rev. Biochem.
71:635-700
3. ^ Rittig MG et al. (2004). "Smooth and rough lipopolysaccharide phenotypes of Brucella induce
different intracellular trafficking and cytokine/chemokine release in human monocytes". Journal of
Leukocyte Biology 5 (4): 196200. DOI:10.1189/jlb.0103015. PMID 12960272.
4. ^ Tsujimoto H et al. (2003). "Diffusion of macrolide antibiotics through the outer membrane of
Moraxella catarrhalis". Journal of Infection and Chemotherapy 74 (4): 10451055.
DOI:10.1007/s101569900025. PMID 11810516.
5. ^ Hershberger C and Binkley SB (1968). "Chemistry and Metabolism of 3-Deoxy-d-mannooctulosonic
Acid. I. STEREOCHEMICAL DETERMINATION". Journal of Biological Chemistry 243 (7): 1578
1584. PMID 4296687.
http://www.jbc.org/cgi/reprint/243/7/1578?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&f
ulltext=3-Deoxy-D-
mannooctulosonic+Acid+&searchid=1&FIRSTINDEX=0&volume=243&issue=7&resourcetype=HW
CIT.
6. ^ Wang, Xiaoyuan and Quinn, Peter J. (2010). "Lipopolysaccharide:Biosynthetic pathway and
structure modification". Progress in Lipid Research 49 (2): 97107.
DOI:10.1016/j.plipres.2009.06.002. PMID 19815028.
7. ^ Ruiz, Natividad; Kahne, Daniel; Silhavy, Thomas J (2009). "Transport of lipopolysaccharide across
the cell envelope: the long road of discovery". Nature Reviews Microbiology 7 (9): 677683.
DOI:10.1038/nrmicro2184. PMC 2790178. PMID 19633680.
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8. ^ Bates J.M. et al. (2007). "Intestinal Alkaline Phosphatase Detoxifies Lipopolysaccharide and
Prevents Inflammation in Response to the Gut Microbiota". Cell Host and Microbe 2 (6): 371382.
DOI:10.1016/j.chom.2007.10.010. PMC 2730374. PMID 18078689.
//www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=2730374.
9. ^ Netea M et al. (2002). "Does the shape of lipid A determine the interaction of LPS with Toll-like
receptors?". Trends Immunol 23 (3): 1359. DOI:10.1016/S1471-4906(01)02169-X. PMID 11864841.
10. ^ Seydel U, Oikawa M, Fukase K, Kusumoto S, Brandenburg K (2000). "Intrinsic conformation of
lipid A is responsible for agonistic and antagonistic activity". Eur J Biochem 267 (10): 30329.
DOI:10.1046/j.1432-1033.2000.01326.x. PMID 10806403.
11. ^ Reeves P, Wang L (2002). "Genomic organization of LPS-specific loci". Curr Top Microbiol
Immunol. Current Topics in Microbiology and Immunology 264 (1): 10935. DOI:10.1007/978-3-642-
56031-6_7. ISBN 978-3-540-42682-0. PMID 12014174.
12. ^ Patil P, Sonti R (2004). "Variation suggestive of horizontal gene transfer at a lipopolysaccharide (lps)
biosynthetic locus in Xanthomonas oryzae pv. oryzae, the bacterial leaf blight pathogen of rice". BMC
Microbiol 4: 40. DOI:10.1186/1471-2180-4-40. PMC 524487. PMID 15473911.
//www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=524487.
13. ^ Warren, HS; Fitting, C; Hoff, E; Adib-Conquy, M; Beasley-Topliffe, L; Tesini, B; Liang, X;
Valentine, C et al. (2010). "Resilience to bacterial infection: difference between species could be due to
proteins in serum". J Infect Dis 201 (2): 223232. DOI:10.1086/649557. PMC 2798011. PMID
20001600. //www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=2798011.
14. ^ Reid RR, Prodeus AP, Khan W, Hsu T, Rosen FS, Carroll MC (1997). "Endotoxin shock in
antibody-deficient mice: unraveling the role of natural antibody and complement in the clearance of
lipopolysaccharide". J. Immunol. 159 (2): 9705. PMID 9218618.
15. ^ Boes M, Prodeus AP, Schmidt T, Carroll MC, Chen J (1998). "A Critical Role of Natural
Immunoglobulin M in Immediate Defense Against Systemic Bacterial Infection". J. Exp. Med. 188
(12): 23816. DOI:10.1084/jem.188.12.2381. PMC 2212438. PMID 9858525.
//www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=2212438.
16. ^ Said EA et al. (2010). "Programmed death-1-induced interleukin-10 production by monocytes
impairs CD4+ T cell activation during HIV infection". Nature Medicine 16 (4): 4529.
DOI:10.1038/nm.2106. PMID 20208540.
17. ^ Poltorak A et al. (1998). "Defective LPS Signaling in C3H/HeJ and C57BL/10ScCr Mice: Mutations
in Tlr4 Gene". Science 282 (5396): 20852088. DOI:10.1126/science.282.5396.2085. PMID 9851930.
18. ^ http://www.nobelprize.org/nobel_prizes/medicine/laureates/2011/press.html

























Lipopolysaccharide (LPS) is the major component of the outer membrane of Gram-negative bacteria.
Lipopolysaccharide is localized in the outer layer of the membrane and is, in noncapsulated strains,
exposed on the cell surface. Within Gram-negative bacteria, the membrane lipopolysaccharides protect the
bacterium against the action of bile salts and lipophilic antibiotics.

Lipopolysaccharides are heat stable endotoxins and have long been recognized as a key factor in septic
shock (septicemia) in humans and, more generally, in inducing a strong immune response in normal
mammalian cells. The lipid A moiety has been identified as critical to the endotoxin activity of
lipopolysaccharide. This was demonstrated by finding identical bioactive results, including endotoxic
activity, between synthetic and natural-sourced E. coli lipid A preparations. The active receptor for
lipopolysaccharide has been identified as the CD14/TLR4/MD2 receptor complex, which promotes the
secretion of proinflammatory cytokines including tumor necrosis factor- and interleukin-1. While the lipid A
component is primarily responsible for immune response activation, the polysaccharide component of
Salmonella enterica LPS is also necessary for NF-B activation.

Lipopolysaccharide preparations have been used in research for the elucidation of LPS structure,
metabolism, immunology, physiology, toxicity, and biosynthesis. They have also been used to induce
synthesis and secretion of growth promoting factors such as interleukins. Because of its connection to
septicemia, lipopolysaccharide has been studied to identify possible targets for antibodies and inhibitors to
LPS biosynthesis.

Lipopolysaccharides can be prepared by extraction from TCA, phenol, or phenol-chloroform-petroleum
ether (for rough strains). TCA extracted lipopolysaccharides are structurally similar to the phenol extracted
ones, with similar electrophoretic patterns and endotoxicity. The primary differences are in the amounts of
nucleic acid and protein contaminants remaining after extraction. The TCA extracts contain ~2% RNA and
~10% denatured proteins, while phenol extracts contain up to 60% RNA and <1% protein. Subsequent
purification by gel filtration chromatography removes much of protein present in the phenol-extracted LPS,
but results in a preparation that contains 10-20% nucleic acids. Further purification using ion exchange
chromatography yields an lipopolysaccharide product which contains <1% protein and <1% RNA.