Inositol phosphate metabolomics: merging genetic perturbation with modernized radiolabeling methods.
Recent discoveries that provide a link between inositol phosphate (IP) signaling and fundamental cellular processes evoke many exciting new hypotheses about IP function, and underscore the importance of understanding how IP synthesis is regulated. Central to studies of IP metabolism is the essential development of efficient, fast, and reproducible methods for quantitative analysis of IPs in systems ranging from simple cell cultures to more complex tissues and whole organisms. Additionally, in many cases there is a need to pharmacologically and/or genetically alter IP kinase and phosphatase activities in order to visualize low abundance inositol signaling messengers. Here, we describe updated methods for rapid analysis of IP metabolism in normal and genetically manipulated Saccharomyces cerevisiae, Arabidopsis thaliana, Drosophila melanogaster, Mus musculus, and Homo sapiens.
Stevenson-Paulik
Jill
J
Department of Pharmacology and Cancer Biology, Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710, USA.
Chiou
Shean-Tai
ST
Frederick
Josh P
JP
dela Cruz
June
J
Seeds
Andrew M
AM
Otto
James C
JC
York
John D
JD
D000818:Animals; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism; D005821:Genetic Techniques; D006801:Humans; D007295:Inositol Phosphates / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*; D007553:Isotope Labeling / Q000379:methods*; D051379:Mice; D007296:Myo-Inositol-1-Phosphate Synthase / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
D007295:Inositol Phosphates; D007296:Myo-Inositol-1-Phosphate Synthase