Mencio Caitlin, Swarup Vimal P, Soliai Marcus, Kuberan Balagurunathan
Interdepartmental Program in Neuroscience, University of Utah, Salt Lake City, UT, USA.
Methods Mol Biol. 2015;1229:37-42. doi: 10.1007/978-1-4939-1714-3_5.
The biological activity of glycosaminoglycans (GAGs) depends greatly on the sulfation pattern present within the GAG chain. Chemical biology of GAGs can be further advanced by preparation of sulfur-isotope-enriched sulfated GAGs. 3'-Phosphoadenosine-5'-phosphosulfate (PAPS) serves as a universal sulfate donor in the sulfation of GAGs by sulfotransferases. Therefore, synthesis of PAPS carrying sulfur isotopes is critical in the preparation of labeled GAGs for biochemical studies. Here we describe a robust in vitro enzymatic synthesis of sulfur isotope-enriched PAPS which allows for heavy- or radio-isotope labeling of GAG chains.
糖胺聚糖（GAGs）的生物活性在很大程度上取决于GAG链中存在的硫酸化模式。通过制备富含硫同位素的硫酸化GAGs，可以进一步推动GAGs的化学生物学研究。3'-磷酸腺苷-5'-磷酸硫酸酯（PAPS）作为硫酸转移酶使GAGs硫酸化过程中的通用硫酸供体。因此，携带硫同位素的PAPS的合成对于制备用于生化研究的标记GAGs至关重要。在此，我们描述了一种强大的体外酶促合成富含硫同位素的PAPS的方法，该方法可实现GAG链的重同位素或放射性同位素标记。
