detection of metabolic fluxes of o and h atoms into intracellular water in mammalian cells检测代谢通量o和h原子的哺乳动物细胞的细胞内的水.pdfVIP

Detection of Metabolic Fluxes of O and H Atoms into Intracellular Water in Mammalian Cells 1 2 3¤a 4 1 Helen W. Kreuzer *, Luca Quaroni *, David W. Podlesak , Theodora Zlateva , Nikki Bollinger , 5¤b 3 5,6 Aaron McAllister , Michael J. Lott , Eric L. Hegg * 1 Pacific Northwest National Laboratory, Richland, Washington, United States of America, 2 Paul Scherrer Institut, Villigen, Switzerland, 3 Department of Biology, University of Utah, Salt Lake City, Utah, United States of America, 4 Saskatchewan Cancer Research Unit and Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada, 5 Department of Chemistry, University of Utah, Salt Lake City, Utah, United States of America, , 6 Department of Biochemistry Molecular Biology, Michigan State University, East Lansing, Michigan, United States of America Abstract Metabolic processes result in the release and exchange of H and O atoms from organic material as well as some inorganic salts and gases. These fluxes of H and O atoms into intracellular water result in an isotopic gradient that can be measured experimentally. Using isotope ratio mass spectroscopy, we revealed that slightly over 50% of the H and O atoms in the intracellular water of exponentially-growing cultured Rat-1 fibroblasts were isotopically distinct from growth medium water. We then employed infrared spectromicroscopy to detect in real time the flux of H atoms in these same cells. Importantly, both of these techniques indicate that the H and O fluxes are dependent on metabolic processes; cells that are in lag phase or are quiescent exhibit a much smaller flux. In addition, water