Updated project metadata. Using an integrated systematic approach to analyse the effects of Hsp90 inhibition in T-cells, we quantified differential changes in the Hsp90-dependent proteome, Hsp90 interactome, and a selection of the transcriptome. Kinetic behaviours in the Hsp90-dependent proteome were assessed using a novel pulse-chase strategy (pulse-chase SILAC, Fierro-Monti et al., accompanying article), detecting effects on both protein stability and synthesis. Global and specific dynamic impacts, including proteostatic responses, are due to direct inhibition of Hsp90 as well as indirect effects. A variety of effects on protein levels and kinetic behaviors are described. Methods : Jurkat cells were treated with DMSO or 1 uM Geldanamycin for 6 or 20h and the proteome was compared to that of untreated cells at both time points. Three biological replicates were prepared and analysed for each condition, one replicate with label inversion. Cell extracts and tryptic digestion were performed according to the FASP protocol. Peptide mixtures were separated into 24 fractions by off-gel isoelectric focusing and analysed by nano-LC-MS/MS on an Orbitrap Velos Instrument. Data were analysed as duplex SILAC using MaxQuant and peptide spectrum matches were filtered at 1% FDR. Output tables were processed further with custom-made perl scripts for further filtering.