INTERLEUKIN-1-BETA PROPEPTIDE IS DETECTED INTRACELLULARLY AND EXTRACELLULARLY WHEN HUMAN MONOCYTES ARE STIMULATED WITH LPS IN-VITRO

被引:25
作者
HIGGINS, GC
FOSTER, JL
POSTLETHWAITE, AE
机构
[1] UNIV TENNESSEE,DEPT BIOCHEM,MEMPHIS,TN 38163
[2] UNIV TENNESSEE,DEPT MED,DIV CONNECT TISSUE DIS,MEMPHIS,TN 38163
[3] VET AFFAIRS MED CTR,MEMPHIS,TN 38163
关键词
D O I
10.1084/jem.180.2.607
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Human interleukin 1 beta (IL-1 beta) is synthesized as an inactive precursor that is cleaved by IL-1 converting enzyme (ICE) between Asp(116) and Ala(117) to form COOH-terminal mature IL-1 beta and NH2-terminal IL-1 beta propeptide. Little is known about the fate of the NH2-terminal cleavage product. In this study, human recombinant (hr)IL-1 beta propeptide (amino acids 2-116) was produced and used to prepare specific antibodies which do not recognize mature human IL-1 beta. These anti-propeptide antibodies were used for immunoprecipitation of biosynthetically labeled proteins from lipopolysaccharide-stimulated human monocytes. Analysis of immunoprecipitates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography revealed that these antibodies recognize precursor IL-1 beta and two unique proteins: one migrating at 17.5 kD and one at 14 kD. The larger of these two proteins has a migration nearly identical to that of the recombinant IL-1 beta propeptide, and most likely represents naturally derived propeptide. The protein migrating at 14 kD may result from a second cleavage by ICE, between Asp(27) and Gly(28). These proteins accumulate intracellularly and extracellularly during pulse-chase experiments, and therefore represent stable products of precursor IL-1 beta cleavage.
引用
收藏
页码:607 / 614
页数:8
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