Chemical approaches for functionally probing the proteome.
With the availability of complete genome sequences, emphasis has shifted toward the understanding of protein function. We have developed a functional proteomic methodology that makes use of chemically reactive fluorescent probes to profile and identify enzymes in complex mixtures by virtue of their catalytic activity. This methodology allows a comparison of changes in activity of multiple enzymes under a variety of conditions using a single two-dimensional separation. The probes can also be used to localize active enzymes in intact cells using fluorescence microscopy. Furthermore, the probes enable screens for selective small molecule inhibitors of each enzyme family member within crude lysates or intact cells. Ultimately, this technology allows the rapid identification of potential drug targets and small molecule lead compounds targeted to them.
Greenbaum
Doron
D
Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143, USA.
Baruch
Amos
A
Hayrapetian
Linda
L
Darula
Zsuzsanna
Z
Burlingame
Alma
A
Medzihradszky
Katlin F
KF
Bogyo
Matthew
M
D000345:Affinity Labels / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002403:Cathepsins / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000378:metabolism*; D007202:Indicators and Reagents; D008099:Liver / Q000378:metabolism; D015336:Molecular Probe Techniques; D015394:Molecular Structure; D010446:Peptide Fragments / Q000737:chemistry; D010447:Peptide Hydrolases / Q000378:metabolism; D019151:Peptide Library; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D012697:Serine Endopeptidases / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
D000345:Affinity Labels; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D010446:Peptide Fragments; D019151:Peptide Library; D020543:Proteome; D002403:Cathepsins; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases