CHARACTERIZATION OF LATENT TRANSFORMING GROWTH-FACTOR-BETA-2 FROM MONKEY KIDNEY-CELLS

被引:15
作者
LIOUBIN, MN
MADISEN, L
ROTH, RA
PURCHIO, AF
机构
[1] ONCOGEN,DEPT MOLEC BIOL,3005 1ST AVE,SEATTLE,WA 98121
[2] STANFORD UNIV,MED CTR,SCH MED,DEPT PHARMACOL,STANFORD,CA 94305
关键词
D O I
10.1210/endo-128-5-2291
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Serum-free medium conditioned by BSC-40 cells was analyzed for the presence of transforming growth factor-beta-2 (TGF-beta-2)-related proteins. Western blot analysis was performed using site-specific antipeptide antibodies directed against the pro- and mature regions of the TGF-beta-2 precursor. When conditioned medium was analyzed by polyacrylamide gel electrophoresis under reducing conditions, proteins with mol wt of 53 kDa (containing both mature and proregion sequences), 34-38 kDa (containing proregion sequences only), and 12 kDa (containing mature sequences) were detected. Under nonreducing conditions, complexes of 60- to 80-kDa, 160- to 200-kDa, as well as 24-kDa mature dimers were seen. Cleavage of mature TGF-beta-2 from its precursor was inhibited by monensin and chloroquin, but not by ammonium chloride or methylamine. Two peaks of bioactivity were detected after fractionation on a TSK column corresponding to mol wt of 130 and 400 kDa. These peaks contained TGF-beta-2 and pro-TGF-beta-2 proteins. Partial purification of the 130-kDa complex followed by N-glyconase digestion indicated that the pro-TGF-beta-2 proteins were glycosylated. These data demonstrate that BSC-40 cells secrete mature TGF-beta-2 complexed with proregion-containing proteins and suggest that this association may contribute to the latency phenomena observed with respect to this growth regulator.
引用
收藏
页码:2291 / 2296
页数:6
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