Isolation and characterization of mutant animal cell line defective in alkyl-dihydroxyacetonephosphate synthase: localization and transport of plasmalogens to post-Golgi compartments.

摘要: We herein isolated plasmalogen-deficient Chinese hamster ovary (CHO) mutant, ZPEG251, with a phenotype of normal import peroxisomal matrix and membrane proteins. In plasmenylethanolamine (PlsEtn) was severely reduced. Complementation analysis by expression genes responsible for the plasmalogen biogenesis suggested that alkyl-dihydroxyacetonephosphate synthase (ADAPS), catalyzing second step biogenesis, deficient in ZPEG251. ADAPS mRNA barely detectable as verified Northern blot reverse transcription-PCR analyses. Defect also assessed immunoblot. As toward delineating functional roles PlsEtn, we investigated its subcellular localization. PlsEtn localized to post-Golgi compartments enriched detergent-resistant membranes. Transport apparently affected lowering cellular ATP, but not inhibitors microtubule assembly vesicular transport. Partitioning cholesterol sphingomyelin, typical feature lipid rafts, impaired cells, including peroxisome assembly-defective mutants, hence suggesting essential lipid-raft architecture CHO cells.