High-yield Expression and Characterization of Syndecan-4 Extracellular, Transmembrane and Cytoplasmic Domains

Bulletin of the Korean chemical society

Jooyoung Song¹  Ji-Sun Kim¹  Sung-Sub Choi¹  Yongae Kim¹ 

关键词: Syndecan;    Syndecan-4;    Transmembrane;    NMR spectroscopy;    Membrane protein;   

DOI  :  

学科分类：化学（综合）

来源: Korean Chemical Society

【 摘 要 】

The syndecan family consists of four transmembrane heparan sulfate proteoglycans present in most cell types and each syndecan shares a common structure containing a heparan sulfate modified extracellular domain, a single transmembrane domain and a C-terminal cytoplasmic domain. To get a better understanding of the mechanism and function of syndecan-4 which is one of the syndecan family, it is crucial to investigate its threedimensional structure. Unfortunately, it is difficult to prepare the peptide because it is membrane-bound protein that transverses the lipid bilayer of the cell membrane. Here, we optimize the expression, purification, and characterization of transmembrane, cytoplasmic and short extracellular domains of syndecan4 (syndecan-4 eTC). Syndecan-4 eTC was successfully obtained with high purity and yield from the M9 medium. The structural information of syndecan-4 eTC was investigated by MALDI-TOF mass (MS) spectrometry, circular dichroism (CD) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. It was confirmed that syndecan-4 eTC had an α-helical multimeric structure like transmembrane domain of syndecan-4 (syndecan- 4 TM) in membrane environments.

【 授权许可】

Unknown