RRC ID 48902
Author Miki T, Awa M, Nishikawa Y, Kiyonaka S, Wakabayashi M, Ishihama Y, Hamachi I.
Title A conditional proteomics approach to identify proteins involved in zinc homeostasis.
Journal Nat Methods
Abstract Zinc signaling and dynamics play significant roles in many physiological responses and diseases. To understand the physiological roles of zinc in detail, comprehensive identification of proteins under high concentration of mobile zinc ion is crucial. We developed a 'conditional proteomics' approach to identify proteins involved in zinc homeostasis based on a chemical proteomic strategy that utilizes designer zinc-responsive labeling reagents to tag such proteins and quantitative mass spectrometry for their identification. We used this method to elucidate zinc dyshomeostasis induced by nitric-oxide-triggered oxidative stress in glioma cells, and we unveiled dynamic changes of the zinc-related proteomes. Moreover, we characterized unknown zinc-rich vesicles generated by oxidative stress as endoplasmic-reticulum- and Golgi-related vesicles.
Volume 13(11)
Pages 931-937
Published 2016-11-1
DOI 10.1038/nmeth.3998
PII nmeth.3998
PMID 27617391
MeSH Binding Sites Blotting, Western Carrier Proteins / genetics Carrier Proteins / metabolism* Cell Line, Tumor Electrophoresis, Polyacrylamide Gel Glioma Green Fluorescent Proteins / genetics HeLa Cells Homeostasis / physiology* Humans Microscopy, Confocal Nitric Oxide / pharmacology Oxidative Stress / drug effects Oxidative Stress / genetics Protein Binding Proteome / genetics Proteome / metabolism* Proteomics / methods* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Staining and Labeling Zinc / metabolism*
IF 30.822
Times Cited 14
DNA material CALR-mCherry-KDEL (RDB15152) PDIA3-mCherry-KDEL (RDB15153) ENPL-mCherry-KDEL (RDB15154) PPIA-mCherry (RDB15155) ARF4-mCherry (RDB15156) mCherry-EF1A1 (RDB15157) mCherry-RAB1A (RDB15158) mCherry-LC3B (RDB15159).