PMID- 17545978
OWN - NLM
STAT- MEDLINE
DCOM- 20070814
LR  - 20161122
IS  - 1750-2799 (Electronic)
IS  - 1750-2799 (Linking)
VI  - 2
IP  - 6
DP  - 2007
TI  - Tandem orthogonal proteolysis-activity-based protein profiling (TOP-ABPP)--a 
      general method for mapping sites of probe modification in proteomes.
PG  - 1414-25
AB  - Activity-based protein profiling (ABPP) utilizes active site-directed chemical 
      probes to monitor the functional state of enzymes directly in native biological 
      systems. Identification of the specific sites of probe labeling on enzymes 
      remains a major challenge in ABPP experiments. In this protocol, we describe an 
      advanced ABPP platform that utilizes a tandem orthogonal proteolysis (TOP) 
      strategy coupled with mass spectrometric analysis to simultaneously identify 
      probe-labeled proteins together with their exact sites of probe modification. 
      Elucidation of probe modification sites reveals fundamental insights into the 
      molecular basis of specific probe-protein interactions. The TOP-ABPP method can 
      be applied to any type of proteomic sample, including those derived from in vitro 
      or in vivo labeling experiments, and is compatible with a variety of chemical 
      probe structures. Completion of the entire protocol, including chemical synthesis 
      of key reagents, requires approximately 8-10 days.
FAU - Weerapana, Eranthie
AU  - Weerapana E
AD  - Department of Cell Biology, The Skaggs Institute for Chemical Biology, 10550 
      North Torrey Pines Road, La Jolla, California 92037, USA.
FAU - Speers, Anna E
AU  - Speers AE
FAU - Cravatt, Benjamin F
AU  - Cravatt BF
LA  - eng
GR  - R01 CA087660/CA/NCI NIH HHS/United States
GR  - R01 CA087660-07/CA/NCI NIH HHS/United States
PT  - Journal Article
PL  - England
TA  - Nat Protoc
JT  - Nature protocols
JID - 101284307
RN  - 0 (Molecular Probes)
RN  - 0 (Proteome)
RN  - 6SO6U10H04 (Biotin)
SB  - IM
MH  - Animals
MH  - Biotin
MH  - Gene Expression Profiling/*methods
MH  - Mice
MH  - *Molecular Probes
MH  - Myocardium/metabolism
MH  - Protein Conformation
MH  - Proteome/*analysis
MH  - Proteomics/methods
EDAT- 2007/06/05 09:00
MHDA- 2007/08/19 09:00
CRDT- 2007/06/05 09:00
PHST- 2007/06/05 09:00 [pubmed]
PHST- 2007/08/19 09:00 [medline]
PHST- 2007/06/05 09:00 [entrez]
AID - nprot.2007.194 [pii]
AID - 10.1038/nprot.2007.194 [doi]
PST - ppublish
SO  - Nat Protoc. 2007;2(6):1414-25. doi: 10.1038/nprot.2007.194.