Reddy U R, Venkatakrishnan G, Maul G G, Roy A K, Ross A H
Department of Neurology, Childrens Hospital of Philadelphia, PA 19104.
Brain Res Mol Brain Res. 1990 Jul;8(2):137-41. doi: 10.1016/0169-328x(90)90058-l.
To facilitate the characterization of nerve growth factor (NGF) receptor and mutated forms of the receptor, we have set up a rapid, efficient transient expression system utilizing COS cells. The human NGF receptor is a 427 amino acid protein with a hydrophobic signal sequence, a 222 amino acid extracellular domain, a single transmembrane domain and a 155 amino acid intracellular domain. The NGF receptor and a truncated form lacking the cytoplasmic domain were expressed in a COS cell expression system. Both recombinant proteins were detected on the cell surface and at a perinuclear site. Specific binding of 125I-NGF to the recombinant proteins was detected by chemical cross-linking. The extracellular domain of the NGF receptor was also expressed in the same system and detected in the COS cell endoplasmic reticulum and in the culture supernatant. This recombinant protein also specifically binds NGF.
为便于对神经生长因子（NGF）受体及其突变形式进行表征，我们利用COS细胞建立了一种快速、高效的瞬时表达系统。人NGF受体是一种由427个氨基酸组成的蛋白质，具有一个疏水信号序列、一个由222个氨基酸组成的细胞外结构域、一个单跨膜结构域和一个由155个氨基酸组成的细胞内结构域。NGF受体和一种缺失细胞质结构域的截短形式在COS细胞表达系统中表达。两种重组蛋白均在细胞表面和核周位点被检测到。通过化学交联检测到125I-NGF与重组蛋白的特异性结合。NGF受体的细胞外结构域也在同一系统中表达，并在COS细胞内质网和培养上清液中被检测到。这种重组蛋白也能特异性结合NGF。
