Extraction and analysis of soluble inositol polyphosphates from yeast

被引：91
作者：
Azevedo, Cristina
Saiardi, Adolfo
机构：
[1] UCL, MRC, Cell Biol Unit, London WC1E 6BT, England
[2] UCL, Dept Biochem & Mol Biol, Mol Cell Biol Lab, London WC1E 6BT, England
来源：
NATURE PROTOCOLS | 2006年 / 1卷 / 05期
基金：
英国医学研究理事会;
关键词：
D O I：
10.1038/nprot.2006.337
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Soluble inositol polyphosphates are implicated in the regulation of many important cellular functions. This protocol to extract and separate inositol polyphosphates from Saccharomyces cerevisiae is divided into three steps: labeling of yeast, extraction of soluble inositol polyphosphates and chromatographic separation. Yeast cells are incubated with tritiated inositol, which is taken up and metabolized into different phosphorylated forms. Soluble inositol polyphosphates are then acid-extracted and fractionated by high-performance liquid chromatography. The radioactivity of each fraction is determined by scintillation counting. This highly sensitive and reproducible method allows the accurate detection of subtle changes in the inositol polyphosphate profile and takes less than 48 h. It can easily be applied to other systems and we have included two adaptations of the protocol, one optimized for mammalian cells and the other for Arabidopsis thaliana.
引用
收藏
页码：2416 / 2422
页数：7
相关论文
共 50 条
  • [21] Regulation of nuclear processes by inositol polyphosphates
    York, John D.
    BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS, 2006, 1761 (5-6): : 552 - 559
  • [22] INOSITOL POLYPHOSPHATES IN SOYBEAN AND COTTON LEAVES
    PHILLIPPY, B
    FASEB JOURNAL, 1993, 7 (04) : A618 - A618
  • [23] INOSITOL POLYPHOSPHATES AND NEURONAL CALCIUM HOMEOSTASIS
    NAHORSKI, SR
    TRENDS IN NEUROSCIENCES, 1988, 11 (10) : 444 - 448
  • [24] Regulation of chromatin remodeling by inositol polyphosphates
    Steger, DJ
    Haswell, ES
    Miller, AL
    Wente, SR
    O'Shea, EK
    SCIENCE, 2003, 299 (5603) : 114 - 116
  • [25] ENANTIOSPECIFIC SYNTHESIS OF INOSITOL POLYPHOSPHATES, PHOSPHATIDYLINOSITIDES, AND ANALOGS FROM (-)-QUINIC ACID
    FALCK, JR
    ABDALI, A
    ACS SYMPOSIUM SERIES, 1991, 463 : 145 - 154
  • [26] On the extraction of inositol from inositol phosphoric acidic salts
    Lindenfeld, K
    BIOCHEMISCHE ZEITSCHRIFT, 1934, 272 : 284 - 289
  • [27] POLYPHOSPHATES OF YEAST CELL VACUOLE
    INDGE, KJ
    JOURNAL OF GENERAL MICROBIOLOGY, 1968, 51 : 447 - &
  • [28] TOTAL SYNTHESIS OF MYO-INOSITOL-POLYPHOSPHATES
    VACCA, JP
    DESOLMS, SJ
    HUFF, JR
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 1987, 194 : 51 - ORGN
  • [29] Membrane-permeant bioactivatable inositol polyphosphates
    Schultz, C
    Rudolf, MT
    Gillandt, H
    TraynorKaplan, AE
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 1997, 214 : 65 - CARB
  • [30] Identity and functions of inorganic and inositol polyphosphates in plants
    Lorenzo-Orts, Laura
    Couto, Daniel
    Hothorn, Michael
    NEW PHYTOLOGIST, 2020, 225 (02) : 637 - 652
12345