小鼠FcγRIIb1和FcγRIIb2胞质区基因的设计、合成、表达及特性分析
Design, synthesis, expression, and characterization of the genes for mouse Fc gamma RIIb1 and Fc gamma RIIb2 cytoplasmic regions.
作者信息
Chen L, Thompson N L, Pielak G J
机构信息
Department of Chemistry, University of North Carolina at Chapel Hill 27599-3290, USA.
出版信息
Protein Sci. 1997 May;6(5):1038-46. doi: 10.1002/pro.5560060510.
The cytoplasmic regions of the mouse low-affinity Fc gamma RII isoforms, mFc gamma RIIb1, and mFc gamma RIIb2, play a key role in signal transduction by mediating different cellular functions. mFc gamma RIIb1 has a 94-residue cytoplasmic region, whereas mFc gamma RIIb2 has a 47-residue cytoplasmic region. Genes encoding the cytoplasmic regions of mFc gamma RIIb1 (b1-94) and mFc gamma RIIb2 (b2-47) were designed, synthesized, and expressed as fusion proteins in Escherichia coli. A sequence-specific protease, thrombin, was used to release the b1-94 peptide, which was purified by using HPLC. The b2-47 peptide was synthesized chemically. CD spectropolarimetry was employed to examine the secondary structures of b1-94 and b2-47. These studies were conducted in aqueous solution, in mixtures of water and trifluoroethanol or methanol, and as a function of temperature. The results indicate that the b1-94 and b2-47 structures are sensitive functions of the solvent environment, and that nonaqueous solvents induce significant alpha-helical structure.
小鼠低亲和力FcγRII同种型mFcγRIIb1和mFcγRIIb2的胞质区域，通过介导不同的细胞功能在信号转导中起关键作用。mFcγRIIb1有一个含94个残基的胞质区域，而mFcγRIIb2有一个含47个残基的胞质区域。编码mFcγRIIb1（b1-94）和mFcγRIIb2（b2-47）胞质区域的基因被设计、合成，并作为融合蛋白在大肠杆菌中表达。一种序列特异性蛋白酶——凝血酶，被用于释放b1-94肽，该肽通过高效液相色谱法纯化。b2-47肽通过化学合成。采用圆二色光谱法检测b1-94和b2-47的二级结构。这些研究在水溶液、水与三氟乙醇或甲醇的混合物中进行，并作为温度的函数。结果表明，b1-94和b2-47的结构是溶剂环境的敏感函数，并且非水溶剂会诱导显著的α-螺旋结构。
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