HEPARAN SULFATE BIOSYNTHETIC FEEDBACKS AND EXTRACELLULAR SULFATASE EXPRESSION

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 Sulf是一个胞外内硫酸酯酶家族，可以修饰细胞表面和细胞外基质中的硫酸乙酰肝素(HS)链。它们在HS链上氨基葡萄糖残基的6-O位释放硫酸盐基团。硫磺酶的活性有助于增强生长因子包括GDNF、BMP、Shh和Wnt的活性，而降低FGF2、HP-EGF、HGF和TGF-β的活性。作为这些活动的结果，根据上下文的不同，Sulf已被观察到作为致癌效应和肿瘤抑制因子。硫磺的活性似乎诱导了生物合成酶的表达变化，导致了表达的HS结构的变化，而这种变化并不一定反映直接的硫磺酶活性。因此，Sulf活性对细胞表型的影响是复杂的。这项工作的目标是(1)确定小鼠胚胎成纤维细胞中HS的结构表型作为HS双糖和寡糖水平上的Sulf酶敲除的函数。这些结果将与对纯化的HS使用重组Sulf酶获得的结果进行比较。 进度更新： 我们发表了两篇关于硫磺基因敲除小鼠精原干细胞巢(1)和肾小球滤过屏障(2)相关的HS结构表型的合作论文。 1.Langsdorf，A.，Schumacher，V.，Shii，X.，Tran，T.，Zaia，J.，Jain，S.，Taglienti，M.，Kreidberg，J.A.，Fine，A.和Ai，X.(2010)Sulff在精原干细胞生态位中的表达调控和功能，糖生物学21,152-161。 2.Schumacher，V.，Schlotzer-Schrehardt，U.S.，Karumachi，S.A.，Shi.，X.，Zaia，J.，Jeruschke，S.，Zhang，D.，Pavenstaedt，H.，Drenckhan，A.，Amann，K.，Ng，C.，Hartwig，S.，Ng，K.-H.，Ho，J.，Kreidberg，J.A.，Taglienti，M.，Royer-Pokora，B.和Ai，X.(2011)WT1对Sulf表达的调控对维持肾小球滤过屏障至关重要，2011年2月22日接受《美国肾脏病学会杂志》。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Sulfs are a family of extracellular endosulfatases that modify heparan sulfate (HS) chains at cell surfaces and in extracellular matrices. They release sulfate groups at the 6-O-position of a subset of glucosamine residues in HS chains. The activity of Sulf enzymes serves to potentiate activities of growth factors including GDNF, BMP, Shh, and Wnt and to reduce activities of FGF2, HP-EGF, HGF, and TGF-¿. As a result of these activities and depending on the context, Sulfs have been observed to serve as oncogenic effectors and as tumor suppressors. Sulf activities appear to induce changes in expression of biosynthetic enzymes, resulting in changes in expressed HS structure that do not necessary reflect the direct Sulf enzymatic activity. As a result, the influences of Sulf activity on cellular phenotype are complex. The goals of this work are to (1) define the HS structural phenotype in mouse embryonic fibroblast cells as a function of Sulf enzyme knockout at both the HS disaccharide and oligosaccharide levels. These results will be compared against those obtained using recombinant Sulf enzymes on purified HS. Progress update: We have published two collaborative papers on the structural phenotypes of HS related to the spermatogonal stem cell niche (1) and the glomerular filtration barrier (2) in Sulf knockout mice. 1. Langsdorf, A., Schumacher, V., Shi, X., Tran, T., Zaia, J., Jain, S., Taglienti, M., Kreidberg, J. A., Fine, A., and Ai, X. (2010) Expression regulation and function of Sulfs in the spermatogonial stem cell niche, Glycobiology 21, 152-161. 2. Schumacher, V., Schlotzer-Schrehardt, U., Karumanchi, S. A., Shi, X., Zaia, J., Jeruschke, S., Zhang, D., Pavenstaedt, H., Drenckhan, A., Amann, K., Ng, C., Hartwig, S., Ng, K.-H., Ho, J., Kreidberg, J. A., Taglienti, M., Royer-Pokora, B., and Ai, X. (2011) WT1 regulation of Sulf expression is crucial to maintaining the glomerular filtration barrier, Journal of the American Society for Nephrology Accepted 2/22/11.