Naider F, Arshava B, Ding F X, Arevalo E, Becker J M
Department of Chemistry, The College of Staten Island of the City University of New York, 10314, USA.
Biopolymers. 2001;60(5):334-50. doi: 10.1002/1097-0282(2001)60:5<334::AID-BIP10175>3.0.CO;2-K.
The alpha-factor tridecapeptide initiates mating in Saccharomyces cerevisiae upon interaction with Ste2p, its cognate G-protein coupled receptor (GPCR). This interaction is being used as a paradigm for understanding the structure and mechanism of activation of GPCRs by medium-sized peptides. In this article, the use of fragments of Ste2p to study its structure is reviewed. Methods of synthesis of peptides corresponding to both extramembranous and transmembrane domains of Ste2p are evaluated and problems that are encountered during synthesis and purification are described. The results from conformational analyses of the peptide fragments using fluorescence spectroscopy, CD, infrared spectroscopy, and NMR spectroscopy in organic-aqueous mixtures and in the presence of detergent micelles and lipid bilayers are critically reviewed. The data obtained to date provide biophysical evidence for the structure of different domains of Ste2p and indicate that peptides corresponding to these domains have unique biophysical tendencies. The studies carried out on Ste2p fragments indicate that valuable information concerning the structure of the intact receptor can be obtained by studying peptide fragments corresponding to domains of these polytopic integral membrane proteins.
α-因子十三肽与它的同源G蛋白偶联受体（GPCR）Ste2p相互作用后，启动酿酒酵母中的交配过程。这种相互作用正被用作理解中-sized肽激活GPCR的结构和机制的范例。在本文中，综述了使用Ste2p片段来研究其结构的情况。评估了对应于Ste2p膜外和跨膜结构域的肽的合成方法，并描述了合成和纯化过程中遇到的问题。对使用荧光光谱、圆二色光谱、红外光谱和核磁共振光谱在有机-水混合物以及存在去污剂胶束和脂质双层的情况下对肽片段进行构象分析的结果进行了严格审查。迄今为止获得的数据为Ste2p不同结构域的结构提供了生物物理证据，并表明对应于这些结构域的肽具有独特的生物物理倾向。对Ste2p片段进行的研究表明，通过研究对应于这些多跨膜整合膜蛋白结构域的肽片段，可以获得有关完整受体结构的有价值信息。
