PMID- 25195567
OWN - NLM
STAT- MEDLINE
DCOM- 20150914
LR  - 20211021
IS  - 1615-9861 (Electronic)
IS  - 1615-9853 (Print)
IS  - 1615-9853 (Linking)
VI  - 15
IP  - 2-3
DP  - 2015 Jan
TI  - Effects of MEK inhibitors GSK1120212 and PD0325901 in vivo using 10-plex 
      quantitative proteomics and phosphoproteomics.
PG  - 462-73
LID - 10.1002/pmic.201400154 [doi]
AB  - Multiplexed isobaric tag based quantitative proteomics and phosphoproteomics 
      strategies can comprehensively analyze drug treatments effects on biological 
      systems. Given the role of mitogen-activated protein/extracellular 
      signal-regulated kinase (MEK) signaling in cancer and mitogen-activated protein 
      kinase (MAPK)-dependent diseases, we sought to determine if this pathway could be 
      inhibited safely by examining the downstream molecular consequences. We used a 
      series of tandem mass tag 10-plex experiments to analyze the effect of two MEK 
      inhibitors (GSK1120212 and PD0325901) on three tissues (kidney, liver, and 
      pancreas) from nine mice. We quantified  approximately  6000 proteins in each tissue, but 
      significant protein-level alterations were minimal with inhibitor treatment. Of 
      particular interest was kidney tissue, as edema is an adverse effect of these 
      inhibitors. From kidney tissue, we enriched phosphopeptides using titanium 
      dioxide (TiO2 ) and quantified 10 562 phosphorylation events. Further analysis by 
      phosphotyrosine peptide immunoprecipitation quantified an additional 592 
      phosphorylation events. Phosphorylation motif analysis revealed that the 
      inhibitors decreased phosphorylation levels of proline-x-serine-proline (PxSP) 
      and serine-proline (SP) sites, consistent with extracellular-signal-regulated 
      kinase (ERK) inhibition. The MEK inhibitors had the greatest decrease on the 
      phosphorylation of two proteins, Barttin and Slc12a3, which have roles in ion 
      transport and fluid balance. Further studies will provide insight into the effect 
      of these MEK inhibitors with respect to edema and other adverse events in mouse 
      models and human patients.
CI  - (c) 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
FAU - Paulo, Joao A
AU  - Paulo JA
AD  - Department of Cell Biology, Harvard Medical School, Boston, MA, USA.
FAU - McAllister, Fiona E
AU  - McAllister FE
FAU - Everley, Robert A
AU  - Everley RA
FAU - Beausoleil, Sean A
AU  - Beausoleil SA
FAU - Banks, Alexander S
AU  - Banks AS
FAU - Gygi, Steven P
AU  - Gygi SP
LA  - eng
GR  - K01 DK093638/DK/NIDDK NIH HHS/United States
GR  - K01 DK098285/DK/NIDDK NIH HHS/United States
GR  - K01DK098285/DK/NIDDK NIH HHS/United States
GR  - K01DK093638/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20141018
PL  - Germany
TA  - Proteomics
JT  - Proteomics
JID - 101092707
RN  - 0 (Benzamides)
RN  - 0 (Phosphopeptides)
RN  - 0 (Protein Kinase Inhibitors)
RN  - 0 (Proteome)
RN  - 0 (Pyridones)
RN  - 0 (Pyrimidinones)
RN  - 33E86K87QN (trametinib)
RN  - 86K0J5AK6M (mirdametinib)
RN  - 9N3CBB0BIQ (Diphenylamine)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
RN  - EC 2.7.12.2 (Mitogen-Activated Protein Kinase Kinases)
SB  - IM
CIN - Proteomics. 2015 Jan;15(2-3):183-4. PMID: 25522341
MH  - Amino Acid Sequence
MH  - Animals
MH  - Benzamides/*pharmacology
MH  - Diphenylamine/*analogs & derivatives/pharmacology
MH  - Humans
MH  - Kidney/drug effects/metabolism
MH  - Liver/drug effects/metabolism
MH  - MAP Kinase Signaling System/drug effects
MH  - Mice
MH  - Mitogen-Activated Protein Kinase Kinases/*antagonists & inhibitors/metabolism
MH  - Mitogen-Activated Protein Kinases/chemistry/metabolism
MH  - Molecular Sequence Data
MH  - Pancreas/drug effects/metabolism
MH  - Phosphopeptides/*analysis/metabolism
MH  - Phosphorylation/drug effects
MH  - Protein Kinase Inhibitors/*pharmacology
MH  - Proteome/chemistry/*metabolism
MH  - Proteomics
MH  - Pyridones/*pharmacology
MH  - Pyrimidinones/*pharmacology
MH  - Tandem Mass Spectrometry
PMC - PMC4515953
MID - NIHMS703175
OTO - NOTNLM
OT  - Barttin
OT  - Cell biology
OT  - GSK1120212
OT  - Multiplexing
OT  - PD0325901
OT  - Phosphoproteomics
COIS- Conflicts of interest The authors acknowledge no conflict of interest.
EDAT- 2014/09/10 06:00
MHDA- 2015/09/15 06:00
PMCR- 2016/01/01
CRDT- 2014/09/09 06:00
PHST- 2014/04/21 00:00 [received]
PHST- 2014/08/05 00:00 [revised]
PHST- 2014/09/01 00:00 [accepted]
PHST- 2014/09/09 06:00 [entrez]
PHST- 2014/09/10 06:00 [pubmed]
PHST- 2015/09/15 06:00 [medline]
PHST- 2016/01/01 00:00 [pmc-release]
AID - 10.1002/pmic.201400154 [doi]
PST - ppublish
SO  - Proteomics. 2015 Jan;15(2-3):462-73. doi: 10.1002/pmic.201400154. Epub 2014 Oct 
      18.