Profiling changes to natively-bound metals during Caenorhabditis elegans development

Relatively little is known about the changing metalloproteome during early development. In this proof-of-concept study, we used size exclusion chromatography-inductively coupled plasma-mass spectrometry (SEC-ICP-MS) to examine the changing soluble metal-binding protein profiles for iron, copper and zinc during the development of the nematode, Caenorhabditis elegans. Samples of eggs, larval stages and young adults were compared using an approach selected to ensure weak metal-ligand bonds were maintained. All three metals showed marked changes in associated proteins and total metal levels per protein mass, and the pattern of this change was unique to each metal. Additionally, to characterise the shifting metabolic needs throughout each life stage we examined changing levels of phosphorus in each developmental stage. The utility of this method can be further exploited through integration with existing proteomics workflows to identify and track the changes in metal-containing proteins during key stages of development.