Substrate Specificity and Domain Functions of Extracellular Heparan Sulfate 6-O-Endosulfatases, QSulf1 and QSulf2

Ai, Xingbin; Do, Anh-Tri; Kusche-Gullberg, Marion; Lindahl, Ulf; Lu, Ke; Emerson, Charles P.

doi:10.1074/jbc.m511902200

Cited by 139 publications

(142 citation statements)

References 24 publications

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“…A portion of the mature heterodimer is secreted, whereas a significant fraction is retained on the cell membrane. It is noteworthy that the QSulfs, in contrast to the HSulfs, are not secreted (28). The Sulfs are highly homologous across species; however, the hydrophilic domains, especially in their central regions, have reduced sequence identity.…”

Section: Discussionmentioning

confidence: 99%

“…This procedure has been established to release QSulfs from intact cells (28). Salt washes of 293 cells transfected with the FLAG/His Sulf plasmids contained both the 75-and 50-kDa bands, as revealed by FLAG and His Western blotting, respectively (Fig.…”

Section: Both Sulf-1 and Sulf-2 Form Disulfide-linked Heterodimers-mentioning

confidence: 99%

“…Thus, in the Sulfs the C-terminal conserved region is separated from the other 8 by insertion of the hydrophilic domain. Ai et al (28) showed that mutations within the hydrophilic domain of QSulf-2 cause a loss of endosulfatase activity against heparan sulfate. We hypothesized that a Sulf mutant, which contains all 9 conserved regions, but not the hydrophilic domain would retain arylsulfatase activity but would lose endosulfatase activity.…”

Section: Both Sulf-1 and Sulf-2 Form Disulfide-linked Heterodimers-mentioning

confidence: 99%

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Functional Consequences of the Subdomain Organization of the Sulfs

Tang

Rosen

2009

Journal of Biological Chemistry

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Sulf-1 and Sulf-2 are novel extracellular sulfatases that act on internal glucosamine 6-O-sulfate modifications within heparan sulfate proteoglycans and regulate their interactions with various signaling molecules, including Wnt ligands. Although the Sulfs are multidomain proteins, there is limited information available about how the subdomains contribute to their enzymatic and signaling activities. In this study, we found that both human Sulfs were synthesized as prepro-enzymes and cleaved by a furin-type proteinase to form disulfide-bond linked heterodimers of 75-and 50-kDa subunits. The mature Sulfs were secreted into conditioned medium, as well as retained on the cell membrane. Although the catalytic center resides in the N-terminal 75-kDa subunit, the C-terminal 50-kDa subunit was indispensable for both arylsufatase and glucosamine 6-O-sulfate-endosulfatase activity. We found that the hydrophilic regions of the Sulfs were essential for endosulfatase activity but not for arylsulfatase activity. Using Edman sequencing, we identified furin-type proteinase cleavage sites in Sulf-1 and Sulf-2. Deletion of these sequences resulted in uncleavable forms of Sulfs. The uncleavable Sulfs retained enzymatic activity. However, they were unable to potentiate Wnt signaling, which may be due to their defective localization into lipid rafts on the plasma membrane.Heparan sulfate proteoglycans (HSPGs) 2 are major components of the extracellular matrix/cell surface and regulate a variety of biological phenomena, including cell proliferation, cell migration, and differentiation (1). These effects are mediated through the ability of HSPGs to bind to a diverse repertoire of protein ligands. Among these are morphogens, growth factors, chemokines, and other classes of molecules (2, 3).HSPGs consist of multiple heparan sulfate (HS) chains covalently linked to a limited set of core proteins. The HS chains contain repeating uronic acid and glucosamine disaccharide units. The binding functions of HSPGs depend on the fine structure of the attached heparan sulfate chains where sulfation modifications occur in four positions (N-, 3-O, and 6-O of glucosamine and 2-O of uronic acid) in highly variegated, yet highly regulated patterns (3, 4). 6-O-Sulfation of glucosamine is established to be critical for certain HSPG functions in organisms from Drosophila through mammals (5, 6).Several years ago, we cloned cDNAs encoding two novel extracellular sulfatases (Sulf-1 and Sulf-2) in human and mouse (7), initiated by the identification of the Sulf-1 ortholog in the quail embryo (QSulf-1) (8). We and others showed that both Sulfs are neutral pH endosulfatases, which remove glucosamine-6-O-sulfate from internal glucosamine residues of highly sulfated subregions within heparin/HSPGs (7, 9, 10). The ability of these enzymes to modulate the heparin/HSPG interactions of a number of growth factors, morphogens, and chemokines has been confirmed in direct binding assays (9, 11-13). In some cellular contexts, the Sulfs act to promote signaling pathways (...

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Section: Discussionmentioning

confidence: 99%

Section: Both Sulf-1 and Sulf-2 Form Disulfide-linked Heterodimers-mentioning

confidence: 99%

Section: Both Sulf-1 and Sulf-2 Form Disulfide-linked Heterodimers-mentioning

confidence: 99%

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Functional Consequences of the Subdomain Organization of the Sulfs

Tang

Rosen

2009

Journal of Biological Chemistry

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“…This backbone is subject to modifications, including de-N-acetylation and N-sulfation of glucosamine, epimerisation of GlcUA resides to iduronic acid (IdoUA), 2-O-sulfation of IdoUA residues, and 6-O-sulfaction and 3-Osulfation of glucosamine residues [3]. Postbiosynthetic processing is carried out by endo-6-sulfatases present at the plasma membrane [4]. The enzymes are both interdependent and incomplete in action, resulting in the typical HS structure where domains of dense sulfation, flanked by regions of intermediate sulfation, are spaced along a nonsulfated backbone [5].…”

Section: Introductionmentioning

confidence: 99%

Specific Glycosaminoglycans Modulate Neural Specification of Mouse Embryonic Stem Cells

et al. 2011

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Mouse embryonic stem (mES) cells express a low sulfated form of heparan sulfate (HS). HS chains displayed by ES cells and their progeny become more complex and more sulfated during progression from pluripotency to neuroectodermal precursors. Sulfated epitopes are important for recognition and binding of a variety of ligands including members of the fibroblast growth factor (FGF) family. We demonstrated previously that mES cells lacking HS cannot undergo neural specification but this activity can be recovered by adding soluble heparin, a highly sulfated glycosaminoglycan (GAG). Therefore, we hypothesized that soluble GAGs might be used to support neural differentiation of HS competent cells and that the mechanisms underlying this activity might provide useful information about the signaling pathways critical for loss of pluripotency and early lineage commitment. In this study, we demonstrate that specific HS/heparin polysaccharides support formation of Sox1 1 neural progenitor cells from wild-type ES cells. This effect is dependent on sulfation pattern, concentration, and length of saccharide. Using a selective inhibitor of FGF signal transduction, we show that heparin modulates signaling events regulating exit from pluripotency and commitment to primitive ectoderm and subsequently neuroectoderm. Interestingly, we were also able to demonstrate that multiple receptor tyrosine kinases were influenced by HS in this system. This suggests roles for additional factors, possibly in cell proliferation or protection from apoptosis, during the process of neural specification. Therefore, we conclude that soluble GAGs or synthetic mimics could be considered as suitable low-cost factors for addition to ES cell differentiation regimes. STEM CELLS 2011;29:629-640 Disclosure of potential conflicts of interest is found at the end of this article.

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“…SULFs are arylendosulfatase which as post synthetic editors selectively liberate 6-O-sulfate groups from heparin sulfates and therefore alter the sulfation patterns of proteoglycans and the binding site of many growth factors [5]. With such unique regulatory activity, SULFs have an important role in many biological processes, such as angiogenesis, cell signalling and embryogenesis [6][7][8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

The association of arylendosulfatase 1 (SULF1) gene polymorphism with recurrent miscarriage

Zahraei

Sheikhha

Kalantar

et al. 2013

J Assist Reprod Genet

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Purpose One of the most common problems in reproductive medicine is recurrent miscarriage (RM). There is increasing evidence showing genetic susceptibility of women is an important risk factor in the occurrence of RM. In recent years, there is a growing interest in sulfate and its role in fetal development. A novel mechanism of SULF1 has been demonstrated for modifying the activities of some growth factors and signalling molecules that have major roles during embryogenesis. The aim of present study was to evaluate the association of SULF1 gene polymorphism (rs6990375 G > A) in Iranian patients with RM. Methods We established a case-control study of 200 Iranian women: 100 patients with the history of two or more RM as cases and 100 healthy women with at least two cases of successful pregnancy and no history of miscarriage as controls. The polymorphism was examined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. Results The genotypic analysis between case and controls showed significant differences (p-value=0.000). Allelic analysis showed no significant correlation (Χ2=3.36, p-value=0.066). The heterozygous genetic variant was significantly higher among healthy women (OR=12.67, 95 % CI=6.47-24.79).Conclusions Our data showed that rs6990375 polymorphism of SULF1 gene could be among one of the factors related to RM in Iranian women. Further evaluation of this polymorphism may be important and need further studies.

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Substrate Specificity and Domain Functions of Extracellular Heparan Sulfate 6-O-Endosulfatases, QSulf1 and QSulf2

Cited by 139 publications

References 24 publications

Functional Consequences of the Subdomain Organization of the Sulfs

Functional Consequences of the Subdomain Organization of the Sulfs

Specific Glycosaminoglycans Modulate Neural Specification of Mouse Embryonic Stem Cells

The association of arylendosulfatase 1 (SULF1) gene polymorphism with recurrent miscarriage

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