Isolation and characterization of a heparin with high anticoagulant activity from the clam Tapes phylippinarum: evidence for the presence of a high content of antithrombin III binding site

Cesaretti, Marina; Luppi, Elisa; Maccari, Francesca; Volpi, Nicola

doi:10.1093/glycob/cwh128

Cited by 61 publications

(55 citation statements)

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“…The significance of high levels of disaccharides with free amino groups in HS from these species is not known, but may signal the existence of an evolutionarily ancient class of biologically important functional motifs. Earlier studies have shown that GAGs are expressed in several other invertebrates (37,47,48). While the earliest diverging animal phylum, Porifera (sponges) appears to express only CS, the expression of both CS and HS is manifest throughout the rest of the animal kingdom (37).…”

Section: Discussionmentioning

confidence: 99%

Evolutionary Differences in Glycosaminoglycan Fine Structure Detected by Quantitative Glycan Reductive Isotope Labeling

Lawrence

Olson

Steele

et al. 2008

Journal of Biological Chemistry

179

195

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To facilitate qualitative and quantitative analysis of glycosaminoglycans, we tagged the reducing end of lyase-generated disaccharides with aniline-containing stable isotopes ( 12 C 6 and 13 C 6 ). Because different isotope tags have no effect on chromatographic retention times but can be discriminated by a mass detector, differentially isotope-tagged samples can be compared simultaneously by liquid chromatography/mass spectrometry and quantified by admixture with known amounts of standards. The technique is adaptable to all types of glycosaminoglycans, and its sensitivity is only limited by the type of mass spectrometer available. We validated the method using commercial heparin and keratan sulfate as well as heparan sulfate isolated from mutant and wild-type Chinese hamster ovary cells, and select tissues from mutant and wild-type mice. This new method provides more robust, reliable, and sensitive means of quantitative evaluation of glycosaminoglycan disaccharide compositions than existing techniques allowing us to compare the chondroitin and heparan sulfate compositions of Hydra vulgaris, Drosophila melanogaster, Caenorhabditis elegans, and mammalian cells. Our results demonstrate significant differences in glycosaminoglycan structure among these organisms that might represent evolutionarily distinct functional motifs.Metazoans make several types of sulfated glycosaminoglycans (GAGs), 2 including keratan sulfate (KS), chondroitin sulfate/dermatan sulfate (CS/DS), and heparan sulfate/heparin (HS). Each type of chain consists of unique disaccharide units. KS consists of galactose (Gal) and GlcNAc ([Gal␤1, 4GlcNAc␤1,3] n ) with variable sulfation at C6 of either sugar. CS/DS assembles as a copolymer of GlcA␤1,3GalNAc␤1,4 and then undergoes various processing reactions, including C5 epimerization of a portion of GlcA units to iduronic acid in DS, O-sulfation at C2 and more rarely at C3 of the uronic acids, and O-sulfation at C4 and C6 of the GalNAc residues (1). HS is the most highly modified GAG, consisting initially of GlcA␤1,4GlcNAc␣1,4 units, which then undergo variable processing by GlcNAc N-deacetylation and N-sulfation, C5 epimerization of some GlcA units to iduronic acid, and O-sulfate addition to C2 of the uronic acids and C6 and more rarely at C3 of the glucosamine units (2). The arrangement of the modified residues along the chain creates binding sites for numerous growth factors, enzymes, and extracellular matrix proteins. The structural variation that can occur makes sulfated GAG chains one of the most complex classes of macromolecules found in nature.GAG fine structure is typically assessed by analyzing the disaccharide composition of an isolated mixture of chains. A number of techniques have been developed to accomplish this task that rely on chemical or enzymatic depolymerization of the chains into their constituent disaccharides, followed by separation via anion exchange chromatography, reversed-phase chromatography with ion pairing agents, or capillary electrophoresis. These techniques separate...

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Section: Discussionmentioning

confidence: 99%

Evolutionary Differences in Glycosaminoglycan Fine Structure Detected by Quantitative Glycan Reductive Isotope Labeling

Lawrence

Olson

Steele

et al. 2008

Journal of Biological Chemistry

179

195

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“…Heparin and heparan sulfate (HS) GAGs bind over 100 different proteins, including enzymes, protease inhibitors, lipoproteins, growth factors, chemokines, selectins, extracellular matrix proteins, receptor proteins, viral coat proteins and nuclear proteins (Linhardt and Toida, 2004;Capila and Linhardt, 2002). Heparin and HS are closely related, overlapping structures that are widely distributed in invertebrates (Medeiros et al, 2000;Nobuo, 2003;Cesaretti et al, 2004), as well as in vertebrates (Gomes and Dietrich, 1982). Moreover, HS GAGs modulate many biological activities including signal transduction (Ibrahimi et al, 2005), inflammation (Peterson et al, 2004), complement activation (Yu et al, 2005) and cAMP-dependent substrate phosphorylation catalyzed by protein kinase (Dittmann et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

“…Thus, it is important to have a detailed understanding of heparin/HS structure to develop structure activity relationships. Much research has been performed to determine the structure of heparin/HS isolated different species and tissues (Gomes and Dietrich, 1982;Medeiros et al, 2000;Nobuo, 2003;Cesaretti et al, 2004). Despite these efforts, there still remains the question about how the fine structure of the GAG moiety of these HSPGs regulates their biological functions.…”

Section: Introductionmentioning

confidence: 99%

Dromedary glycosaminoglycans: Molecular characterization of camel lung and liver heparan sulfate

Warda

Linhardt

2006

Comparative Biochemistry and Physiology Part B: Biochemistry an

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Glycosaminoglycans (GAGs) are the portion of a proteoglycan that determine its final shape and function. The molecular structure of predominant GAG species in camel liver and lung is reported for the first time. The one-humped camel survives in an extreme, arid habitat and, thus, offers a good model to study the role of glycomics on homeostasis. Heparan sulfate (HS) from the lung and liver of the one-humped camel were isolated. Characterization of these newly isolated glycosaminoglycans included 1 H NMR spectroscopy and disaccharide compositional analysis. The relative molecular weight of these GAGs was estimated by gradient polyacrylamide gel electrophoresis and their degree of sulfation was also assessed. Anticoagulant activity was determined using an anti-factor Xa assay and the HS from camel lung shows ~50% of heparin's activity. The structural differences of camel liver GAGs compared to human and porcine liver heparin and HS is discussed. Camel lung heparan sulfate resembles both heparin and HS in its structure and properties suggesting that it is either a highly sulfated form of HS, a mixture of heparin and HS or an undersulfated heparin.

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“…The proteoglycans chondroitin sulfate and heparan sulfate, as well as the structurally related heparin (31), have been observed in a wide variety of animals, ranging from mammals (3-6, 32, 33), to birds (34), clams (35), fruit flies, and worms (24). Nothing, however, is known about these molecules in insects of medical importance, where they may play a role in the transmission of pathogens.…”

Section: Discussionmentioning

confidence: 99%

Mosquito Heparan Sulfate and Its Potential Role in Malaria Infection and Transmission

Sinnis

Coppi

Toida

et al. 2007

Journal of Biological Chemistry

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Heparan sulfate has been isolated for the first time from the mosquito Anopheles stephensi, a known vector for Plasmodium parasites, the causative agents of malaria. Chondroitin sulfate, but not dermatan sulfate or hyaluronan, was also present in the mosquito. The glycosaminoglycans were isolated, from salivary glands and midguts of the mosquito in quantities sufficient for disaccharide microanalysis. Both of these organs are invaded at different stages of the Plasmodium life cycle. Mosquito heparan sulfate was found to contain the critical trisulfated disaccharide sequence, 34)␤-D-GlcNS6S(1 3 4)-␣-L-IdoA2S(13, that is commonly found in human liver heparan sulfate, which serves as the receptor for apolipoprotein E and is also believed to be responsible for binding to the circumsporozoite protein found on the surface of the Plasmodium sporozoite. The heparan sulfate isolated from the whole mosquito binds to circumsporozoite protein, suggesting a role within the mosquito for infection and transmission of the Plasmodium parasite.Malaria infection is initiated with the bite of an infected anophelene mosquito, which injects Plasmodium sporozoites into the skin of the vertebrate host (Fig. 1A). Sporozoites migrate from the site of injection, enter the blood stream, and are carried to the liver, where they invade hepatocytes and develop into exoerythrocytic forms. Many lines of evidence suggest that clearance of sporozoites by the liver is mediated by the binding of the sporozoite major surface protein, the circumsporozoite protein (CSP), 2 to heparan sulfate proteoglycans in the liver (1, 2). Liver heparan sulfate is more highly sulfated than heparan sulfate from other organs (3-6) and frequently contains the fully biosynthetically modified, highly sulfated sequence: 34)-GlcNS6S(1 3 4)IdoA2S(13 (Fig. 2). Such fully modified sequences, while commonly found in the mast cell glycosaminoglycan, heparin, are infrequently found in heparan sulfate. Some heparan sulfates, such as those present in the liver, have a richer content of fully modified sequences in their high sulfate domains. These sequences have been shown to bind to apoE and facilitate the clearance of lipoprotein remnants from the blood by the liver (7,8). ApoE containing remnant lipoproteins inhibits Plasmodium sporozoite infectivity and decreases clearance of CSP from the blood circulation, suggesting that sporozoites bind to the same hepatic heparan sulfate proteoglycans as remnant lipoproteins (9). This hypothesis is further supported by the finding that CSP binds to the same decasaccharide structure that binds to apoE (10, 11).Less is known about the role of proteoglycans in the mosquito host, where the parasite encounters a variety of mosquito matrices as it develops (Fig. 1B). When the mosquito takes a blood meal from an infected vertebrate host, gametocytes present in the blood meal differentiate, and fertilization occurs. The zygote develops into an ookinete, which crosses the midgut wall, stops at the basal lamina of the midgut, and develops into an oo...

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Isolation and characterization of a heparin with high anticoagulant activity from the clam Tapes phylippinarum: evidence for the presence of a high content of antithrombin III binding site

Cited by 61 publications

References 35 publications

Evolutionary Differences in Glycosaminoglycan Fine Structure Detected by Quantitative Glycan Reductive Isotope Labeling

Evolutionary Differences in Glycosaminoglycan Fine Structure Detected by Quantitative Glycan Reductive Isotope Labeling

Dromedary glycosaminoglycans: Molecular characterization of camel lung and liver heparan sulfate

Mosquito Heparan Sulfate and Its Potential Role in Malaria Infection and Transmission

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