A Generic Method for Expression and Use of “Tagged” Soluble Versions of Cell Surface Receptors
作者： Erik A. Whitehorn , Emily Tate , Stephen D. Yanofsky , Lynn Kochersperger , Ann Davis
DOI: 10.1038/NBT1195-1215
关键词: Chinese hamster ovary cell 、 Fusion protein 、 Membrane protein 、 Chimeric gene 、 Signal peptide 、 Cell surface receptor 、 Biochemistry 、 Epitope 、 Receptor 、 Biology 、 Biotechnology 、 Molecular medicine 、 Applied Microbiology and Biotechnology 、 Bioengineering 、 Biomedical engineering
摘要: A general method for expression, purification, immobilization, detection and radiolabeling of extracellular domains (ECD) type I membrane proteins. The interleukin-1 receptor (IL-lRtI), the α-subunit interleukin-2 (IL-2Rα) E-selectin are used as illustrative examples cell surface receptors. DNA encoding ECD proteins fused at their 3′ end to a chimeric which serves generically “tag” recombinant ECD. resulting fusion protein contains substrate sequence kinase-A (PKA) adjacent signal from human placental alkaline phosphatase (HPAP). HPAP directs formation phosphatidylinositol-glycan (PI-G) anchorage surface. When these genes expressed in CHO cells, ECDs detected on can be released by treatment with phosphatidylinos-itol-specific phospholipase-C (PI-PLC). Based processing known occur native HPAP, twenty amino acids remain C-terminus high affinity monoclonal antibody was generated against this common epitope. This detect, purify immobilize ECDs. In addition, radiolabeled 32P PKA maintain ability bind natural ligands. “tagging” has been successfully applied many other serve
nature.com
sci-hub.se 参考文章(25)
M Fountoulakis, J F Juranville, D Stüber, E K Weibel, G Garotta, Purification and biochemical characterization of a soluble human interferon gamma receptor expressed in Escherichia coli. Journal of Biological Chemistry. ,vol. 265, pp. 13268- 13275 ,(1990) , 10.1016/S0021-9258(19)38294-8
G Fuh, M G Mulkerrin, S Bass, N McFarland, M Brochier, J H Bourell, D R Light, J A Wells, The human growth hormone receptor. Secretion from Escherichia coli and disulfide bonding pattern of the extracellular binding domain. Journal of Biological Chemistry. ,vol. 265, pp. 3111- 3115 ,(1990) , 10.1016/S0021-9258(19)39741-8
Y Takebe, M Seiki, J Fujisawa, P Hoy, K Yokota, K Arai, M Yoshida, N Arai, SR alpha promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat. Molecular and Cellular Biology. ,vol. 8, pp. 466- 472 ,(1988) , 10.1128/MCB.8.1.466
S Rose-John, P C Heinrich, Soluble receptors for cytokines and growth factors: generation and biological function Biochemical Journal. ,vol. 300, pp. 281- 290 ,(1994) , 10.1042/BJ3000281
M G Low, Biochemistry of the glycosyl-phosphatidylinositol membrane protein anchors. Biochemical Journal. ,vol. 244, pp. 1- 13 ,(1987) , 10.1042/BJ2440001
V C Gibbs, S R Williams, P W Gray, R D Schreiber, D Pennica, G Rice, D V Goeddel, The extracellular domain of the human interferon gamma receptor interacts with a species-specific signal transducer. Molecular and Cellular Biology. ,vol. 11, pp. 5860- 5866 ,(1991) , 10.1128/MCB.11.12.5860
Warren J. Leonard, Joel M. Depper, Gerald R. Crabtree, Stuart Rudikoff, Janet Pumphrey, Richard J. Robb, Martin Krönke, Penny B. Svetlik, Nancy J. Peffer, Thomas A. Waldmann, Warner C. Greene, Molecular cloning and expression of cDNAs for the human interleukin-2 receptor Nature. ,vol. 311, pp. 626- 631 ,(1984) , 10.1038/311626A0
B. L. Li, J. A. Langer, B. Schwartz, S. Pestka, Creation of phosphorylation sites in proteins: construction of a phosphorylatable human interferon alpha Proceedings of the National Academy of Sciences of the United States of America. ,vol. 86, pp. 558- 562 ,(1989) , 10.1073/PNAS.86.2.558
R. Micanovic, L. D. Gerber, J. Berger, K. Kodukula, S. Udenfriend, Selectivity of the cleavage/attachment site of phosphatidylinositol-glycan-anchored membrane proteins determined by site-specific mutagenesis at Asp-484 of placental alkaline phosphatase Proceedings of the National Academy of Sciences of the United States of America. ,vol. 87, pp. 157- 161 ,(1990) , 10.1073/PNAS.87.1.157
Stephen P. Eisenberg, Ron J. Evans, William P. Arend, Evie Verderber, Michael T Brewer, Charles H. Hannum, Robert C. Thompson, Primary structure and functional expression from complementary DNA of a human interleukin-1 receptor antagonist Nature. ,vol. 343, pp. 341- 346 ,(1990) , 10.1038/343341A0