PMID- 25271054
OWN - NLM
STAT- MEDLINE
DCOM- 20151026
LR  - 20211021
IS  - 1535-3907 (Electronic)
IS  - 1535-3893 (Print)
IS  - 1535-3893 (Linking)
VI  - 13
IP  - 12
DP  - 2014 Dec 5
TI  - In-depth quantitative proteomic analysis of de novo protein synthesis induced by 
      brain-derived neurotrophic factor.
PG  - 5707-14
LID - 10.1021/pr5006982 [doi]
AB  - Measuring the synthesis of new proteins in the context of a much greater number 
      of pre-existing proteins can be difficult. To overcome this obstacle, 
      bioorthogonal noncanonical amino acid tagging (BONCAT) can be combined with 
      stable isotope labeling by amino acid in cell culture (SILAC) for comparative 
      proteomic analysis of de novo protein synthesis (BONLAC). In the present study, 
      we show that alkyne resin-based isolation of l-azidohomoalanine (AHA)-labeled 
      proteins using azide/alkyne cycloaddition minimizes contamination from 
      pre-existing proteins. Using this approach, we isolated and identified 7414 
      BONCAT-labeled proteins. The nascent proteome isolated by BONCAT was very similar 
      to the steady-state proteome, although transcription factors were highly enriched 
      by BONCAT. About 30% of the methionine residues were replaced by AHA in our 
      BONCAT samples, which allowed for identification of methionine-containing 
      peptides. There was no bias against low-methionine proteins by BONCAT at the 
      proteome level. When we applied the BONLAC approach to screen for brain-derived 
      neurotrophic factor (BDNF)-induced protein synthesis, 53 proteins were found to 
      be significantly changed 2 h after BDNF stimulation. Our study demonstrated that 
      the newly synthesized proteome, even after a short period of stimulation, can be 
      efficiently isolated by BONCAT and analyzed to a depth that is similar to that of 
      the steady-state proteome.
FAU - Zhang, Guoan
AU  - Zhang G
AD  - Department of Biochemistry and Molecular Pharmacology, double daggerDepartments of Cell 
      Biology, Physiology, and Neuroscience and Psychiatry, Kimmel Center for Biology 
      and Medicine at the Skirball Institute, New York University School of Medicine , 
      New York, New York 10016, United States.
FAU - Bowling, Heather
AU  - Bowling H
FAU - Hom, Nancy
AU  - Hom N
FAU - Kirshenbaum, Kent
AU  - Kirshenbaum K
FAU - Klann, Eric
AU  - Klann E
FAU - Chao, Moses V
AU  - Chao MV
FAU - Neubert, Thomas A
AU  - Neubert TA
LA  - eng
GR  - P01 HD023315/HD/NICHD NIH HHS/United States
GR  - T32 MH019524/MH/NIMH NIH HHS/United States
GR  - R01 NS047384/NS/NINDS NIH HHS/United States
GR  - R37 NS034007/NS/NINDS NIH HHS/United States
GR  - R01 HD23315/HD/NICHD NIH HHS/United States
GR  - R01 AG025970/AG/NIA NIH HHS/United States
GR  - R01 NS21072/NS/NINDS NIH HHS/United States
GR  - R01 NS034007/NS/NINDS NIH HHS/United States
GR  - R01 NS021072/NS/NINDS NIH HHS/United States
GR  - P30 NS050276/NS/NINDS NIH HHS/United States
GR  - T32 MH019524-20/MH/NIMH NIH HHS/United States
GR  - S10 RR027990/RR/NCRR NIH HHS/United States
GR  - R56 NS021072/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20141013
PL  - United States
TA  - J Proteome Res
JT  - Journal of proteome research
JID - 101128775
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Proteome)
RN  - 0 (azidohomoalanine)
RN  - OF5P57N2ZX (Alanine)
SB  - IM
MH  - Alanine/analogs & derivatives/chemistry
MH  - Brain-Derived Neurotrophic Factor/*pharmacology
MH  - Chromatography, Liquid
MH  - HEK293 Cells
MH  - Humans
MH  - Mass Spectrometry
MH  - Protein Biosynthesis/*drug effects
MH  - Proteome/*analysis/chemistry/metabolism
MH  - Proteomics/*methods
MH  - Reproducibility of Results
PMC - PMC4261974
OTO - NOTNLM
OT  - BDNF
OT  - BONCAT
OT  - mass spectrometry
OT  - proteomics
OT  - pulsed SILAC
OT  - translation
EDAT- 2014/10/02 06:00
MHDA- 2015/10/27 06:00
PMCR- 2015/10/01
CRDT- 2014/10/02 06:00
PHST- 2014/10/02 06:00 [entrez]
PHST- 2014/10/02 06:00 [pubmed]
PHST- 2015/10/27 06:00 [medline]
PHST- 2015/10/01 00:00 [pmc-release]
AID - 10.1021/pr5006982 [doi]
PST - ppublish
SO  - J Proteome Res. 2014 Dec 5;13(12):5707-14. doi: 10.1021/pr5006982. Epub 2014 Oct 
      13.