CHARACTERIZATION OF RECOMBINANT EXTRACELLULAR DOMAIN OF HUMAN INTERLEUKIN-10 RECEPTOR

被引:54
作者
TAN, JC [1 ]
BRAUN, S [1 ]
RONG, H [1 ]
DIGIACOMO, R [1 ]
DOLPHIN, E [1 ]
BALDWIN, S [1 ]
NARULA, SK [1 ]
ZAVODNY, PJ [1 ]
CHOU, CC [1 ]
机构
[1] SCHERING PLOUGH CORP,RES INST,DEPT IMMUNOL,KENILWORTH,NJ 07033
关键词
D O I
10.1074/jbc.270.21.12906
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The extracellular region of the human interleukin-10 (hIL-10) receptor was expressed using a myeloma cell line and was purified to homogeneity by Ligand-affinity chromatography. SDS-polyacrylamide gel electrophoresis analysis indicated that the soluble receptor is glycosylated and has an apparent molecular mass of 35,000-45,000. Under native conditions, soluble hIL-10 receptor was determined by gel filtration to be a monomeric protein. Soluble hIL-10 receptor was able to inhibit the binding of I-125-hTL-10 to the full-length receptor and was able to antagonize the effect of human IL-10 in cell proliferation and cytokine synthesis inhibition. The apparent dissociation constant (K-d) of soluble hIL-10 receptor was determined to be 563 +/- 59 pM, approximately 2- to 10-fold higher than that found on intact cells (Tan, J. C., Idelicato, S. R., Narula, S. IL, Zavodny, P. J., and Chou, C.-C. (1993) J. Biol. Chem. 268, 21053-21059; Liu, Y., Wei, S. H.-Y., Ho, A S.-Y., de Waal Malefyt, R., and Moore, IL m. (1994) J. Immunol. 152, 1821-1829). When hIL-10 binds soluble hIL-10 receptor in solution, a single complex was detected by gel filtration, and the complex was found to consist of two hIL-10 dimers and four soluble receptor monomers, suggesting that hIL-10 may induce a novel mode of oligomerization of the receptor upon binding.
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页码:12906 / 12911
页数:6
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