A new chemical probe for phosphatidylinositol kinase activity.
Phosphatidylinositol kinases (PIKs) are key enzymatic regulators of membrane phospholipids and membrane environments that control many aspects of cellular function, from signal transduction to secretion, through the Golgi apparatus. Here, we have developed a photoreactive "clickable" probe, PIK-BPyne, to report the activity of PIKs. We investigated the selectivity and efficiency of the probe to both inhibit and label PIKs, and we compared PIK-BPyne to a wortmannin activity-based probe also known to target PIKs. We found that PIK-BPyne can act as an effective in situ activity-based probe, and for the first time, report changes in PI4K-IIIβ activity induced by the hepatitis C virus. These results establish the utility of PIK-BPyne for activity-based protein profiling studies of PIK function in native biological systems.
Life Sciences Division, National Research Council of Canada, 100 Sussex Drive, Ottawa, K1A 0R6 (Canada).
Allison R
AR
Sherratt
Neda
N
Nasheri
Craig S
CS
McKay
Shifawn
S
O'Hara
Ashley
A
Hunt
Zhibin
Z
Ning
Daniel
D
Figeys
Natalie K
NK
Goto
John Paul
JP
Pezacki
D000480:Alkynes / Q000737:chemistry / Q000494:pharmacology*; D001577:Benzophenones / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D005456:Fluorescent Dyes / Q000737:chemistry / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D015394:Molecular Structure; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013329:Structure-Activity Relationship
D000480:Alkynes; D001577:Benzophenones; D005456:Fluorescent Dyes; C047723:benzophenone; D017853:Phosphotransferases (Alcohol Group Acceptor)
HCV; activity-based protein profiling; enzyme activity; phosphatidylinositol kinases; photo-crosslinking