Profiling the specific reactivity of the proteome with non-directed activity-based probes.
BACKGROUND
The field of proteomics aims to characterize dynamics in protein function on a global level. However, several classes of proteins, in particular low abundance proteins, remain difficult to characterize using standard proteomics technologies. Recently, chemical strategies have emerged that profile classes of proteins based on activity rather than quantity, thereby greatly facilitating the analysis of low abundance constituents of the proteome.


RESULTS
In order to expand the classes of proteins susceptible to analysis by activity-based methods, we have synthesized a library of biotinylated sulfonate esters and applied its members to complex proteomes under conditions that distinguish patterns of specific protein reactivity. Individual sulfonates exhibited unique profiles of proteome reactivity that in extreme cases appeared nearly orthogonal to one another. A robustly labeled protein was identified as a class I aldehyde dehydrogenase and shown to be irreversibly inhibited by members of the sulfonate library.


CONCLUSIONS
Through screening the proteome with a non-directed library of chemical probes, diverse patterns of protein reactivity were uncovered. These probes labeled protein targets based on properties other than abundance, circumventing one of the major challenges facing contemporary proteomics research. Considering further that the probes were found to inhibit a target enzyme's catalytic activity, the methods described herein should facilitate the identification of compounds possessing both selective proteome reactivities and novel bioactivities.
Adam
G C
GC
The Skaggs Institute for Chemical Biology, La Jolla, CA 92037, USA.
Cravatt
B F
BF
Sorensen
E J
EJ
D000444:Aldehyde Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000737:chemistry; D015195:Drug Design; D066298:In Vitro Techniques; D008297:Male; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry; D008969:Molecular Sequence Data; D020543:Proteome / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013451:Sulfonic Acids / Q000138:chemical synthesis / Q000737:chemistry
D015335:Molecular Probes; D020543:Proteome; D011994:Recombinant Proteins; D013451:Sulfonic Acids; D001710:Biotin; D000444:Aldehyde Dehydrogenase