Purification of cachectin, a lipoprotein lipase-suppressing hormone secreted by endotoxin-induced RAW 264.7 cells

B. Beutler, J. Mahoney, N. Le Trang, P. Pekala, A. Cerami

Research output: Contribution to journalArticle

568 Citations (Scopus)

Abstract

Previous studies have indicated that endotoxin and other bacterial and protozoal products can stimulate macrophages to produce a factor that can suppress the activity of the enzyme lipoprotein lipase (LPL), in vivo and in vitro. In the present report we describe the purification of this factor, cachectin, to apparent homogeneity from the conditioned medium of endotoxin-stimulated RAW 264.7 cells. The isolated protein has an isoelectric point of 4.7 and a subunit molecular weight of 17,000. Although cachectin's isoelectric point and molecular weight are similar to those described for interleukin 1, pure cachectin has no leukocyte-activating factor (LAF) activity. Cachectin at a concentration of 10-11 M has the ability to suppress the LPL activity of the 3T3-L1 adipocyte cell line by 80%. Binding studies using radiolabeled cachectin and 3T3-L1 adipocytes and C2 myotubules revealed ~104 high-affinity receptors per cell on both cell types (K(a), 3 x 109). Cachectin receptors were also present on liver membranes but were absent on erythrocytes and lymphocytes. The isolation of cachectin and characterization of its receptor should facilitate further investigations into the role of cachectin and other macrophage mediators in the metabolic derangements that occur during infection and cachexia.

Original languageEnglish (US)
Pages (from-to)984-995
Number of pages12
JournalJournal of Experimental Medicine
Volume161
Issue number5
DOIs
StatePublished - 1985

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Lipoprotein Lipase
Endotoxins
Tumor Necrosis Factor-alpha
Hormones
Isoelectric Point
Adipocytes
Molecular Weight
Macrophages
3T3-L1 Cells
Cachexia
Tumor Necrosis Factor Receptors
Conditioned Culture Medium
RAW 264.7 Cells
Interleukin-1
Leukocytes
Erythrocytes
Lymphocytes
Cell Line
Membranes
Liver

ASJC Scopus subject areas

  • Immunology

Cite this

Purification of cachectin, a lipoprotein lipase-suppressing hormone secreted by endotoxin-induced RAW 264.7 cells. / Beutler, B.; Mahoney, J.; Le Trang, N.; Pekala, P.; Cerami, A.

In: Journal of Experimental Medicine, Vol. 161, No. 5, 1985, p. 984-995.

Research output: Contribution to journalArticle

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AU - Cerami, A.

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