Center for Biotechnology and Interdisciplinary Studies, Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
Department of Biochemistry, College of Medicine, Konyang University, Konyang Hospital, Daejeon 302-718, Korea.
Biosensors (Basel). 2015 Jul 23;5(3):500-12. doi: 10.3390/bios5030500.
Tissue inhibitor of metalloproteinases-3 (TIMP-3) belongs to a family of proteins that regulate the activity of matrix metalloproteinases (MMPs), which can process various bioactive molecules such as cell surface receptors, chemokines, and cytokines. Glycosaminoglycans (GAGs) interact with a number of proteins, thereby playing an essential role in the regulation of many physiological/patho-physiological processes. Both GAGs and TIMP/MMPs play a major role in many cell biological processes, including cell proliferation, migration, differentiation, angiogenesis, apoptosis, and host defense. In this report, a heparin biosensor was used to map the interaction between TIMP-3 and heparin and other GAGs by surface plasmon resonance spectroscopy. These studies show that TIMP-3 is a heparin-binding protein with an affinity of ~59 nM. Competition surface plasmon resonance analysis indicates that the interaction between TIMP-3 and heparin is chain-length dependent, and N-sulfo and 6-O-sulfo groups (rather than the 2-O-sulfo groups) in heparin are important in the interaction of heparin with TIMP-3. Other GAGs (including chondroitin sulfate (CS) type E (CS-E)and CS type B (CS-B)demonstrated strong binding to TIMP-3, while heparan sulfate (HS), CS type A (CSA), CS type C (CSC), and CS type D (CSD) displayed only weak binding affinity.
组织金属蛋白酶抑制剂 3（TIMP-3）属于一类能够调节基质金属蛋白酶（MMPs）活性的蛋白质家族，MMPs 能够加工多种生物活性分子，如细胞表面受体、趋化因子和细胞因子。糖胺聚糖（GAGs）与许多蛋白质相互作用，从而在许多生理/病理生理过程的调节中发挥重要作用。GAGs 和 TIMP/MMPs 在许多细胞生物学过程中都发挥着重要作用，包括细胞增殖、迁移、分化、血管生成、凋亡和宿主防御。在本报告中，使用肝素生物传感器通过表面等离子体共振光谱法来绘制 TIMP-3 与肝素和其他 GAGs 之间的相互作用图。这些研究表明，TIMP-3 是一种肝素结合蛋白，亲和力约为 59 nM。竞争表面等离子体共振分析表明，TIMP-3 与肝素之间的相互作用依赖于链长，肝素中的 N-磺酸基和 6-O-磺酸基（而不是 2-O-磺酸基）对于肝素与 TIMP-3 的相互作用很重要。其他 GAGs（包括硫酸软骨素 E（CS-E）和硫酸软骨素 B（CS-B））与 TIMP-3 表现出强烈的结合，而硫酸乙酰肝素（HS）、硫酸软骨素 A（CSA）、硫酸软骨素 C（CSC）和硫酸软骨素 D（CSD）仅显示出较弱的结合亲和力。
