Dynamic changes of the Caenorhabditis elegans proteome during ontogenesis assessed by quantitative analysis with 15N metabolic labeling.
The development of the nematode Caenorhabditis elegans is a highly dynamic process. Although various studies have assessed global transcriptome changes, information on the dynamics of the proteome during ontogenesis is not available. We metabolically labeled C. elegans by using ¹⁵N ammonium chloride as a precursor in Escherichia coli feeding bacteria grown in minimal media as a new cost-effective technique. Quantitative proteome analysis was performed by LC-MS/MS in animals harvested at different times during ontogenesis. We identified and quantified 245 proteins at all larval stages in two independent replicates. Between larval stages (20 and 40 h after hatching) 61 were found to change significantly in level. Among those ribosomal proteins, aminoacyl tRNA synthetases and enzymes of energy metabolism increased in abundance, while extracellular matrix proteins and muscle proteins dominated groups displaying reduced levels. Moreover, changes observed for selected proteins such as VIT-6 and SOD-1 matched with previously published findings confirming the validity of our approach. The metabolic labeling technique applied seems well suited to assess changes in the proteome changes of C. elegans in a quantitative manner during larval development. The data set generated provides the basis for further exploitation of the role of individual proteins or protein clusters during ontogenesis.
ZIEL Research Center of Nutrition and Food Sciences, Abteilung Biochemie, Technische Universität München, Gregor-Mendel-Strasse 2, 85350 Freising, Germany. kersting@tum.de
Kerstin E
KE
Geillinger
Katja
K
Kuhlmann
Martin
M
Eisenacher
Helmut E
HE
Meyer
Hannelore
H
Daniel
Britta
B
Spanier
D000818:Animals; D017173:Caenorhabditis elegans / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D016000:Cluster Analysis; D007553:Isotope Labeling / Q000379:methods; D007814:Larva; D009587:Nitrogen Isotopes / Q000032:analysis* / Q000378:metabolism; D060066:Protein Interaction Maps; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D012269:Ribosomal Proteins / Q000032:analysis / Q000378:metabolism; D053719:Tandem Mass Spectrometry
D009587:Nitrogen Isotopes; D020543:Proteome; D012269:Ribosomal Proteins