RRC ID 1886
Author Harada K, Fukusaki E, Bamba T, Sato F, Kobayashi A.
Title In vivo 15N-enrichment of metabolites in suspension cultured cells and its application to metabolomics.
Journal Biotechnol. Prog.
Abstract The incorporation of stable isotopes in suspension cultured cells is very simple and useful as a preliminary experimental method in the experimental scene of plant metabolomics to elucidate the metabolic profiles of mutants and transformants. Stable isotope methods would afford a dynamic explanation of turnover speed that would concern the metabolic flux. Utilization of suspension cultured cells allows genes to be easily induced or suppressed, culture conditions to be controlled, and samples to be easily prepared. Stable isotope tracing allows an index of metabolic flux to be obtained. Here we present an experiment feeding (15)N-labeled inorganic salts to Arabidopsis (cell line T87) and Coptis cultured cells. Results of a comparison of (15)N labeling ratios of amino acids derived from T87 cells cultured under light with those cultured in the dark corresponded to transcriptional expressions revealed by microarray experiments published previously, demonstrating the validity of this procedure. Furthermore, (15)N labeling ratios of Coptis cultured cells revealed arginine and lysine metabolism inhibition, which should result in inhibition of polyamine biosynthesis and cell division. This very simple experiment allowed us to uncover metabolic dynamic features of the plant cell. Therefore this method is very useful for forming working hypotheses and experimental design.
Volume 22(4)
Pages 1003-11
Published 2006-7
DOI 10.1021/bp060139z
PMID 16889377
MeSH Amino Acids / antagonists & inhibitors Amino Acids / chemistry Amino Acids / metabolism Arabidopsis / chemistry Arabidopsis / cytology Arabidopsis / metabolism* Cell Culture Techniques / methods Cell Division Cell Line Chorismate Mutase / analysis Chorismate Mutase / biosynthesis Chorismate Mutase / genetics Coptis / chemistry Coptis / cytology Coptis / metabolism* Mass Spectrometry / methods Nitrates / chemistry Nitrates / metabolism* Nitrates / pharmacology Nitrogen Isotopes Protein Array Analysis / methods Quaternary Ammonium Compounds / chemistry Quaternary Ammonium Compounds / metabolism* Quaternary Ammonium Compounds / pharmacology Sensitivity and Specificity
IF 1.947
Times Cited 26
Arabidopsis / Cultured plant cells, genes rpc00008