Substrate specificity of 6-O-endosulfatase (Sulf-2) and its implications in synthesizing anticoagulant heparan sulfate

Pempe, Elizabeth H.; Burch, Tanya C.; Law, Courtney J; Liu, Jian

doi:10.1093/glycob/cws092

Cited by 26 publications

(19 citation statements)

References 44 publications

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“…Enzymatic activity of the preparations was assessed using 4-MUS as a pseudosubstrate (25), and HS chains as a natural substrate followed by disaccharide analysis ( Supplemental Table S1). Results showed that HSulf-2 essentially targets [uronic acid-(2S)-N-sulfoglucosamine-(6S)] ([UA(2S)GlcNS(6S)]) trisulfated disaccharides, along with a small but significant decrease of the [⌬UA-GlcNS(6S)] disulfated disaccharide species, in agreement with previously published data (25)(26)(27)(28). To study in detail this desulfation mechanism without being hindered by the natural structural complexity of the intact polysaccharide, we analyzed HSulf-2 activity on a highly sulfated oligosaccharide that mimics heparin structure as well as the inner regions of HS S-domains.…”

Section: Analysis Of Hsulf-2 Enzymatic Activity On a Heparin Oligosacsupporting

confidence: 90%

HSulf sulfatases catalyze processive and oriented 6‐O‐desulfation of heparan sulfate that differentially regulates fibroblast growth factor activity

et al. 2013

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Sulfs are extracellular sulfatases that have emerged recently as critical regulators of heparan sulfate (HS) activities through their ability to catalyze specific 6-O-desulfation of the polysaccharide. Consequently, Sulfs have been involved in many physiological and pathological processes, and notably for Sulf-2, in the development of cancers with poor prognosis. Despite growing interest, little is known about the structure and activity of these enzymes and the way they induce dynamic remodeling of HS 6-O-sulfation status. Here, we have combined an array of analytical approaches, including mass spectrometry, NMR, HS oligosaccharide sequencing, and FACS, to dissect HSulf-2 sulfatase activity, either on a purified octasaccharide used as a mimic of HS functional domains, or on intact cell-surface HS chains. In parallel, we have studied the functional consequences of HSulf-2 activity on fibroblast growth factor (FGF)-induced mitogenesis and found that the enzyme could differentially regulate FGF1 and FGF2 activities. Notably, these data supported the existence of precise 6-O-sulfation patterns for FGF activation and provided new insights into the saccharide structures involved. Altogether, our data bring to light an original processive enzymatic mechanism, by which HSulfs catalyze oriented alteration of HS 6-O-desulfation patterns and direct fine and differential regulation of HS functions.

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Section: Analysis Of Hsulf-2 Enzymatic Activity On a Heparin Oligosacsupporting

confidence: 90%

HSulf sulfatases catalyze processive and oriented 6‐O‐desulfation of heparan sulfate that differentially regulates fibroblast growth factor activity

et al. 2013

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“…6-OST isoform 1 knockout is lethal to mice at the late embryonic stage, displaying systematic skeletal growth retardation, aberrant lung morphology, and reduced placental angiogenesis, whereas 6-OST isoform 2 knockout mice appear to be normal and fertile 20 . The cellular concentration of 6- O -sulfation is influenced by expression levels of 6-OST and endoglucosamine-6-sulfatases which selectively hydrolyze the 6- O -sulfo group present in the highly sulfated domains of HS containing the disaccharide repeating unit -IdoA2S-GlcNS6S- 21 .…”

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confidence: 99%

Structure Based Substrate Specificity Analysis of Heparan Sulfate 6-O-Sulfotransferases

Moon

et al. 2016

ACS Chem. Biol.

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Heparan sulfate (HS) is a sulfated polysaccharide exhibiting essential physiological functions. HS 6-O-sulfotransferase (6-OST) transfers a sulfo group to the 6-OH position of glucosamine units to confer a variety of HS biological activities. There are three different isoforms of 6-OST in the human genome. Here, we report crystal structures of the ternary complex of 6-OST with the sulfo donor analog 3′-phosphoadenosine 5′-phosphate and three different oligosaccharide substrates at 1.95 to 2.1 Å resolutions. Structural and mutational analyses reveal amino acid residues that contribute to catalysis and substrate recognition of 6-OST. Unexpectedly, the structures reveal 6-OST engages HS in a completely different orientation than other HS sulfotransferases, and sheds light on the basic HS requirements for specificity. These findings also contribute structural information to understand mutations in human 6-OST isoform 1 associated with the human genetic disease idiopathic hypogonadotrophic hypogonadism characterized by delayed or lack of puberty. Keywords oligosaccharides; heparin; sulfated carbohydrates; crystal structure Heparan sulfates (HS) are sulfated polysaccharides that are found on the surface of mammalian cells attached to core syndecan and glypican proteins and are secreted into the extracellular matrix, associated with perlecan, agrin and collagen XVIII 1, 2 . HS contains a disaccharide repeating unit of glucuronic acid (GlcA) or iduronic acid (IdoA) and glucosamine (GlcN). Both IdoA and GlcN are capable of carrying sulfo groups, while GlcA can be sulfated to a lesser extent. HS play important roles in a wide range of physiological HHS Public AccessAuthor manuscript ACS Chem Biol. Author manuscript; available in PMC 2018 January 20. Author Manuscript Author ManuscriptAuthor Manuscript Author Manuscript and pathophysiological functions including assisting in viral/bacterial infection, inflammatory responses, blood coagulation, angiogenesis, and embryonic development 3 . Notably, HS is implicated in the formation of amyloid plaques contributing to Alzheimer's disease 4 . HS carry out their wide range of functions through interactions with proteins such as growth factors, protease inhibitors, proteases, cytokines, chemokines and morphogens 1, 2, 5 . Many of these interactions rely on distinctive sulfated saccharide sequences in HS for high specificity.The HS 6-O-sulfotransferase isoforms (6-OSTs) are members the HS biosynthetic enzyme family that transfer a sulfo group from 3′-phosphoadenosine 5′-phosphosulfate (PAPS) to the 6-OH of GlcN. HS biosynthesis occurs in the Golgi and endoplasm reticulum and involves glycosyltransferases, an epimerase and several sulfotransferases. A backbone polysaccharide with a disaccharide repeating unit of GlcA and N-acetyl glucosamine (-GlcA-GlcNAc-) is initially synthesized by HS polymerase. The backbone polysaccharide then undergoes a series of modifications ( Supplementary Figure 1) 6 . In general, modification begins with N-deacetylation/N-sulfation by the N-dea...

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“…It is first well established that HS 6-O-sulfation is a major structural determinant for the interaction with many chemokines, including CXCL12 (78-80), CXCL8 (81,82), CXCL4 (83,84) and CCL5 (85). For the latter, the resolution of the CCL5 crystal structure in complex with heparin provided further evidence of the contribution of 6-O-S group in the interaction (85).…”

Section: Discussion: Potential Roles Of the Sulfs During Inflammationmentioning

confidence: 95%

HS and Inflammation: A Potential Playground for the Sulfs?

et al. 2020

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Heparan sulfate (HS) is a complex polysaccharide abundantly found in extracellular matrices and cell surfaces. HS participates in major cellular processes, through its ability to bind and modulate a wide array of signaling proteins. HS/ligand interactions involve saccharide domains of specific sulfation pattern. Assembly of such domains is orchestrated by a complex biosynthesis machinery and their structure is further regulated at the cell surface by post-synthetic modifying enzymes. Amongst them, extracellular sulfatases of the Sulf family catalyze the selective removal of 6-O-sulfate groups, which participate in the binding of many proteins. As such, increasing interest arose on the regulation of HS biological properties by the Sulfs. However, studies of the Sulfs have so far been essentially restricted to the fields of development and tumor progression. The aim of this review is to survey recent data of the literature on the still poorly documented role of the Sulfs during inflammation, and to widen the perspectives for the study of this intriguing regulatory mechanism toward new physiopathological processes.

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Substrate specificity of 6-O-endosulfatase (Sulf-2) and its implications in synthesizing anticoagulant heparan sulfate

Cited by 26 publications

References 44 publications

HSulf sulfatases catalyze processive and oriented 6‐O‐desulfation of heparan sulfate that differentially regulates fibroblast growth factor activity

HSulf sulfatases catalyze processive and oriented 6‐O‐desulfation of heparan sulfate that differentially regulates fibroblast growth factor activity

Structure Based Substrate Specificity Analysis of Heparan Sulfate 6-O-Sulfotransferases

HS and Inflammation: A Potential Playground for the Sulfs?

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