Assessing enzyme activities using stable isotope labeling and mass spectrometry.
Activity-based protein profiling has emerged as a valuable technology for labeling, enriching, and assessing protein activities from complex mixtures. This is primarily accomplished via a two-step identification and quantification process. Here we show a highly quantitative and streamlined method, termed catch-and-release activity profiling of enzymes (CAPE), which reduces this procedure to a single step. Furthermore the CAPE approach has the ability to detect small quantitative changes that may have been missed by alternative mass spectrometry-based techniques.
Everley
Patrick A
PA
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Gartner
Carlos A
CA
Haas
Wilhelm
W
Saghatelian
Alan
A
Elias
Joshua E
JE
Cravatt
Benjamin F
BF
Zetter
Bruce R
BR
Gygi
Steven P
SP
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D045744:Cell Line, Tumor; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism; D063065:Organophosphonates / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism*; D014362:Trypsinogen / Q000378:metabolism*
D009363:Neoplasm Proteins; D063065:Organophosphonates; D010455:Peptides; D014362:Trypsinogen; D014357:Trypsin