GDP-capture compound--a novel tool for the profiling of GTPases in pro- and eukaryotes by capture compound mass spectrometry (CCMS).
The functional isolation of proteome subsets based on small molecule-protein interactions is an increasingly popular and promising field in functional proteomics. Entire protein families may be profiled on the basis of their common interaction with a metabolite or small molecule inhibitor. This is enabled by novel multifunctional small molecule probes. One platform approach in this field are Capture Compounds that contain a small molecule of interest to bind target proteins, a photo-activatable reactivity function to covalently trap bound proteins, and a sorting function to isolate Capture Compound-protein conjugates from complex biological samples for direct trypsinisation and protein identification by liquid chromatography/mass spectrometry (CCMS). We here present the synthesis and application of a novel GDP-Capture Compound for the functional enrichment of GTPases, a pivotal protein family that exerts key functions in signal transduction. We present data from CCMS experiments on two biological lysates from Escherichia coli and from human-derived Hek293 cells. The GDP-Capture Compound robustly captures a wide range of different GTPases from both systems and will be a valuable tool for the proteomic profiling of this important protein family.
Luo
Yan
Y
Caprotec Bioanalytics GmbH, Volmerstrasse 5, 12489 Berlin, Germany.
Fischer
Jenny J
JJ
Baessler
Olivia Y Graebner Neé
OY
Schrey
Anna K
AK
Ungewiss
Jan
J
Glinski
Mirko
M
Sefkow
Michael
M
Dreger
Mathias
M
Koester
Hubert
H
D002457:Cell Extracts; D002460:Cell Line; D002853:Chromatography, Liquid; D030562:Databases, Protein; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005057:Eukaryotic Cells / Q000201:enzymology*; D020558:GTP Phosphohydrolases / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry; D011387:Prokaryotic Cells / Q000201:enzymology*; D040901:Proteomics; D014357:Trypsin / Q000494:pharmacology
D002457:Cell Extracts; D029968:Escherichia coli Proteins; D006153:Guanosine Diphosphate; D014357:Trypsin; D020558:GTP Phosphohydrolases