Mutants in a macrophage-like cell line are defective in plasmalogen biosynthesis, but contain functional peroxisomes

Raphael A. Zoeller, Shanthi Rangaswamy, Haya Herscovitz, William B Rizzo, Amiya K. Hajra, Arun K. Das, Hugo W. Moser, Ann Moser, Paul B. Lazarow, Manuel J. Santos

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Abstract

We have used a fluorescence-activated cytotoxicity protocol, 9-(1′-pyrene)nonanol (P9OH)/UV selection (Morand, O. H., Allen, L.-A. H., Zoeller, R. A., and Raetz, C. R. H. (1990) Biochim. Biophya. Acta 1034, 132-141), to isolate a series of plasmalogen-deficient mutants in a murine, macrophage-like cell line, RAW 264.7. Three of these mutants, RAW.7, RAW.12, and RAW.108, displayed varying degrees of plasmalogen deficiency (48, 17, and 14% of wild-type levels, respectively), and all three mutants were deficient in peroxisomal dihydroxyacetone phosphate (DHAP) acyltransferase activity (5% of wild-type). Unlike previously described Chinese hamster ovary (CHO) cell mutants, the RAW mutants contained intact, functional, peroxisomes and normal levels of alkyl-DHAP synthase activity, a peroxisomal, membrane-bound enzyme. In RAW.7 and RAW.108 cells, the loss of peroxisomal DHAP acyltransferase is the primary lesion. RAW.12 displayed not only a deficiency in the DHAP acyltransferase activity, but also displayed a second lesion in the biosynthetic pathway, a deficiency in Δ1′-desaturase activity (plasmanylethanolamine desaturase, EC 1.14.99.19), the final step in plasmenylethanolamine biosynthesis. The deficiencies expressed in the mutants represent unique lesions in plasmalogen biosynthesis. Since the RAW cell line is a macrophage-like responsive cell line, these mutants can be used to examine the role of plasmalogens in cellular functions such as arachidonic acid metabolism, prostaglandin synthesis, protein secretion, and signal transduction.

Original languageEnglish (US)
Pages (from-to)8299-8306
Number of pages8
JournalJournal of Biological Chemistry
Volume267
Issue number12
StatePublished - Apr 25 1992

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glycerone-phosphate O-acyltransferase
Plasmalogens
Peroxisomes
plasmanylethanolamine desaturase
Macrophages
Biosynthesis
Cells
Cell Line
Signal transduction
Biosynthetic Pathways
Cytotoxicity
Cricetulus
Arachidonic Acid
Metabolism
Prostaglandins
Ovary
Signal Transduction
Fluorescence
Membranes
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Zoeller, R. A., Rangaswamy, S., Herscovitz, H., Rizzo, W. B., Hajra, A. K., Das, A. K., ... Santos, M. J. (1992). Mutants in a macrophage-like cell line are defective in plasmalogen biosynthesis, but contain functional peroxisomes. Journal of Biological Chemistry, 267(12), 8299-8306.

Mutants in a macrophage-like cell line are defective in plasmalogen biosynthesis, but contain functional peroxisomes. / Zoeller, Raphael A.; Rangaswamy, Shanthi; Herscovitz, Haya; Rizzo, William B; Hajra, Amiya K.; Das, Arun K.; Moser, Hugo W.; Moser, Ann; Lazarow, Paul B.; Santos, Manuel J.

In: Journal of Biological Chemistry, Vol. 267, No. 12, 25.04.1992, p. 8299-8306.

Research output: Contribution to journalArticle

Zoeller, RA, Rangaswamy, S, Herscovitz, H, Rizzo, WB, Hajra, AK, Das, AK, Moser, HW, Moser, A, Lazarow, PB & Santos, MJ 1992, 'Mutants in a macrophage-like cell line are defective in plasmalogen biosynthesis, but contain functional peroxisomes', Journal of Biological Chemistry, vol. 267, no. 12, pp. 8299-8306.
Zoeller, Raphael A. ; Rangaswamy, Shanthi ; Herscovitz, Haya ; Rizzo, William B ; Hajra, Amiya K. ; Das, Arun K. ; Moser, Hugo W. ; Moser, Ann ; Lazarow, Paul B. ; Santos, Manuel J. / Mutants in a macrophage-like cell line are defective in plasmalogen biosynthesis, but contain functional peroxisomes. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 12. pp. 8299-8306.
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abstract = "We have used a fluorescence-activated cytotoxicity protocol, 9-(1′-pyrene)nonanol (P9OH)/UV selection (Morand, O. H., Allen, L.-A. H., Zoeller, R. A., and Raetz, C. R. H. (1990) Biochim. Biophya. Acta 1034, 132-141), to isolate a series of plasmalogen-deficient mutants in a murine, macrophage-like cell line, RAW 264.7. Three of these mutants, RAW.7, RAW.12, and RAW.108, displayed varying degrees of plasmalogen deficiency (48, 17, and 14{\%} of wild-type levels, respectively), and all three mutants were deficient in peroxisomal dihydroxyacetone phosphate (DHAP) acyltransferase activity (5{\%} of wild-type). Unlike previously described Chinese hamster ovary (CHO) cell mutants, the RAW mutants contained intact, functional, peroxisomes and normal levels of alkyl-DHAP synthase activity, a peroxisomal, membrane-bound enzyme. In RAW.7 and RAW.108 cells, the loss of peroxisomal DHAP acyltransferase is the primary lesion. RAW.12 displayed not only a deficiency in the DHAP acyltransferase activity, but also displayed a second lesion in the biosynthetic pathway, a deficiency in Δ1′-desaturase activity (plasmanylethanolamine desaturase, EC 1.14.99.19), the final step in plasmenylethanolamine biosynthesis. The deficiencies expressed in the mutants represent unique lesions in plasmalogen biosynthesis. Since the RAW cell line is a macrophage-like responsive cell line, these mutants can be used to examine the role of plasmalogens in cellular functions such as arachidonic acid metabolism, prostaglandin synthesis, protein secretion, and signal transduction.",
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AU - Rangaswamy, Shanthi

AU - Herscovitz, Haya

AU - Rizzo, William B

AU - Hajra, Amiya K.

AU - Das, Arun K.

AU - Moser, Hugo W.

AU - Moser, Ann

AU - Lazarow, Paul B.

AU - Santos, Manuel J.

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