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Biochemical Characterization of Interleukin 1 From a Human Monocytic Cell Line
Author(s) -
Krakauer Teresa
Publication year - 1985
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.37.5.511
Subject(s) - thp1 cell line , monocytic leukemia , iodoacetamide , concanavalin a , isoelectric focusing , biology , cell culture , microbiology and biotechnology , phenylglyoxal , hl60 , biochemistry , lymphokine , monocyte , arginine , cell , in vitro , immunology , amino acid , enzyme , cysteine , genetics
A protein with interleukin 1 (IL‐1) activity was obtained from the human acute monocytic leukemia cell line (THP‐1) and purified by ultrafiltration, Ultrogel AcA54 chromatography, isoelectric focusing, and discontinuous polyacrylam‐ ide gel electrophoresis. Like the pi 6.8 species of IL‐1 from human peripheral blood monocytes (PBM), the cell line IL‐1 has a molecular weight (Mw) of 14,000, a pL of 6.8, is heat labile, and does not bind to concanavalin A‐ Sepharose. Chemical modification of arginine residues by phenylglyoxal or sulfhydryl groups by N‐ethyl maleimide and iodoacetamide completely destroys the activity of IL‐1 from THP‐1 cells as well as that of the pl 6.8 component from PBM. In contrast, the pi 5.1 component of PBM IL‐1 is resistant to heat denaturation and sulfhydryl reagents, although it is totally inactivated by phenylglyoxal. IL‐1 from THP‐1 cells enhanced the proliferative response of the same subpopulations of PNA‐ thymocytes as IL‐1 from PBM. These observations suggest that IL‐1 derived from this cell line is similar to the IL‐1 pi 6.8 species produced by human monocytes, and distinct from the pi 5.1 species.