期刊
PROCESS BIOCHEMISTRY
卷 46, 期 5, 页码 1166-1171出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2011.02.006
关键词
Syndecan-4; Transmembrane protein; Expression; Purification; NMR
资金
- Ministry of Education, Science and Technology [2010-0021954]
- HUFS
- National Research Foundation of Korea [2010-0021954] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
The transmembrane heparan sulfate proteoglycan, syndecan-4, plays potentially important roles that may influence tissue development and repair as well as the pathogenesis of numerous diseases. Recently, a number of studies have been conducted on the structural properties and biological functions of the syndecan-4 cytoplasmic domain or extracellular domain. However, previous attempts to understand the structure and function of the syndecan-4 transmembrane domain (Syd4-TM) have been marred by experimental adversities due to insufficient yields and low solubility. To investigate the structural and functional properties of Syd4-TM, an efficient method of preparing milligram quantities of the corresponding peptide is necessary. Herein, we report an efficient method for the recombinant expression and purification of Syd4-TM peptide. Peptide was released from the fusion protein, and then purified by semi-preparative reversed-phase HPLC. Finally, more than 5 mg of Syd4-TM peptide was obtained with high purity from 11 of M9 minimal media under optimized conditions. The preliminary biophysical properties of recombinant Syd4-TM peptide were studied by circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy. The analysis of the CD spectrum shows that Syd4-TM adopts a stable a-helical structure in micelle environments. And solution NMR studies showed that Syd4-TM forms an asymmetric dimer in micelles. (C) 2011 Elsevier Ltd. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据