Detection of Glycosaminoglycans by Polyacrylamide Gel Electrophoresis and Silver Staining.
Detection of Glycosaminoglycans by Polyacrylamide Gel Electrophoresis and Silver Staining.
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DOI：
10.3791/62319
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发表时间：
2021-02-25
影响因子：
1.2
通讯作者：
Schmidt, Eric P.
中科院分区：
文献类型：
--
作者：
LaRiviere, Wells B.;Han, Xiaorui;Oshima, Kaori;McMurtry, Sarah A.;Linhardt, Robert J.;Schmidt, Eric P.
硫酸化糖胺聚糖（GAG），如硫酸乙酰肝素（HS）和硫酸软骨素（CS），是多糖，其通过与蛋白质的静电相互作用对组织结构做出贡献并影响信号传导过程。GAG聚合物的特定长度对于它们对同源配体的结合亲和力特别重要。本报告描述了一种从生物样品中分离和纯化GAG的技术，并使用聚丙烯酰胺凝胶电泳来近似它们的大小。硫酸化糖胺聚糖（GAG）如硫酸乙酰肝素（HS）和硫酸软骨素（CS）普遍存在于活生物体中，并且在各种基本生物结构和过程中起关键作用。作为聚合物，GAG以多分散混合物的形式存在，所述多分散混合物含有多糖链，其范围可以从4000 Da到远远超过40，000 Da。在这些链内存在硫酸化结构域，赋予负电荷模式，从而促进与同源蛋白质配体的带正电荷残基的相互作用。糖胺聚糖的硫酸化结构域必须具有足够的长度以允许这些静电相互作用。为了了解GAG在生物组织中的功能，研究者必须能够分离、纯化和测量GAG的大小。本报告描述了一种实用和多功能的聚丙烯酰胺凝胶电泳技术，可用于解决从各种生物组织类型中分离的GAG之间相对较小的大小差异。
Sulfated glycosaminoglycans (GAGs), such as heparan sulfate (HS) and chondroitin sulfate (CS), are polysaccharides that both contribute to tissue structure and influence signaling processes via electrostatic interaction with proteins. The specific length of GAG polymers is of particular importance to their binding affinity for cognate ligands. This report describes a technique to isolate and purify GAGs from biological samples and use polyacrylamide gel electrophoresis to approximate their size. Sulfated glycosaminoglycans (GAGs) such as heparan sulfate (HS) and chondroitin sulfate (CS) are ubiquitous in living organisms and play a critical role in in a variety of basic biological structures and processes. As polymers, GAGs exist as a polydisperse mixture containing polysaccharide chains that can range from 4000 Da to well over 40,000 Da. Within these chains exists domains of sulfation, conferring a pattern of negative charge that facilitates interaction with positively charged residues of cognate protein ligands. Sulfated domains of GAGs must be of sufficient length to allow for these electrostatic interactions. To understand the function of GAGs in biological tissues, the investigator must be able to isolate, purify, and measure the size of GAGs. This report describes a practical and versatile polyacrylamide gel electrophoresis-based technique that can be leveraged to resolve relatively small differences in size between GAGs isolated from a variety of biological tissue types.
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影响因子：
15.9
作者：
MANKIN, HJ;LIPPIELL.L
通讯作者：
LIPPIELL.L
DOI：
10.1165/rcmb.2017-0428oc
发表时间：
2018-09-01
影响因子：
6.4
作者：
Haeger, Sarah M.;Liu, Xinyue;Schmidt, Eric P.
通讯作者：
Schmidt, Eric P.
影响因子：
2.9
作者：
JAY, GD;CULP, DJ;JAHNKE, MR
通讯作者：
JAHNKE, MR
影响因子：
82.9
作者：
Schmidt, Eric P.;Yang, Yimu;Janssen, William J.;Gandjeva, Aneta;Perez, Mario J.;Barthel, Lea;Zemans, Rachel L.;Bowman, Joel C.;Koyanagi, Dan E.;Yunt, Zulma X.;Smith, Lynelle P.;Cheng, Sara S.;Overdier, Katherine H.;Thompson, Kathy R.;Geraci, Mark W.;Douglas, Ivor S.;Pearse, David B.;Tuder, Rubin M.
通讯作者：
Tuder, Rubin M.
影响因子：
4.8
作者：
Pempe, Elizabeth H.;Xu, Yongmei;Harris, Edward N.
通讯作者：
Harris, Edward N.