Previous structural analyses of diphosphoinositol polyphosphates in biological systems have relied largely on NMR analysis. For example, in Dictyostelium discoideum, diphosphoinositol pentakisphosphate was determined by NMR to be 4- and/or 6-PPInsP5, and the bisdiphosphoinositol tetrakisphosphate was found to be 4,5-bisPPInsP4 and/or 5,6-bisPPInsP4 [Laussmann, Eujen, Weisshuhn, Thiel and Vogel (1996) Biochem. J. 315, 715-720]. We now describe three recent technical developments to aid the analysis of these compounds, not just in Dictyostelium, but also in a wider range of biological systems: (i) improved resolution and sensitivity of detection of PPInsP5 isomers by microbore metal-dye-detection HPLC; (ii) the use of the enantiomerically specific properties of a rat hepatic diphosphatase; (iii) chemical synthesis of enantiomerically pure reference standards of all six possible PPInsP5 isomers. Thus we now demonstrate that the major PPInsP5 isomer in Dictyostelium is 6-PPInsP5. Similar findings obtained using the same synthetic standards have been published [Laussmann, Reddy, Reddy, Falck and Vogel (1997) Biochem. J. 322, 31-33]. In addition, we show that 10-25% of the Dictyostelium PPInsP5 pool is comprised of 5-PPInsP5. The biological significance of this new observation was reinforced by our demonstration that 5-PPInsP5 is the predominant PPInsP5 isomer in four different mammalian cell lines (FTC human thyroid cancer cells, Swiss 3T3 fibroblasts, Jurkat T-cells and Chinese hamster ovary cells). The fact that the cellular spectrum of diphosphoinositol polyphosphates varies across phylogenetic boundaries underscores the value of our technological developments for future determinations of the structures of this class of compounds in other systems.
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Research Article|
October 15 1997
Biological variability in the structures of diphosphoinositol polyphosphates in Dictyostelium discoideum and mammalian cells
Claudia ALBERT;
Claudia ALBERT
*Universitäts-Krankenhaus Eppendorf, Institut für Physiologische Chemie, Abt. für Enzymchemie, Martinistr. 52, D-20246 Hamburg, Germany
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T. Stephen SAFRANY;
T. Stephen SAFRANY
†Inositol Lipid Section, Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, U.S.A.
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E. Michael BEMBENEK;
E. Michael BEMBENEK
‡Medical Products Division, E.I. DuPont De Nemours and Co., 549 Albany Street, Boston MA 02118, U.S.A.
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Komandla Malla REDDY;
Komandla Malla REDDY
§Departments of Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75235-9038, U.S.A.
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K. Kishta REDDY;
K. Kishta REDDY
§Departments of Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75235-9038, U.S.A.
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J. R. FALCK;
J. R. FALCK
§Departments of Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75235-9038, U.S.A.
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Martina BRÖCKER;
Martina BRÖCKER
ǁUniversity Hospital Bergmannsheil, Ruhr-University Bochum, Endocrinology Laboratory, House 10, D44789 Bochum, Germany
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B. Stephen SHEARS;
B. Stephen SHEARS
1
†Inositol Lipid Section, Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, U.S.A.
1To whom correspondence should be addressed.
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W. Georg MAYR
W. Georg MAYR
*Universitäts-Krankenhaus Eppendorf, Institut für Physiologische Chemie, Abt. für Enzymchemie, Martinistr. 52, D-20246 Hamburg, Germany
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Publisher: Portland Press Ltd
Received:
October 14 1996
Revision Received:
May 27 1997
Accepted:
June 12 1997
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1997
1997
Biochem J (1997) 327 (2): 553–560.
Article history
Received:
October 14 1996
Revision Received:
May 27 1997
Accepted:
June 12 1997
Citation
Claudia ALBERT, T. Stephen SAFRANY, E. Michael BEMBENEK, Komandla Malla REDDY, K. Kishta REDDY, J. R. FALCK, Martina BRÖCKER, B. Stephen SHEARS, W. Georg MAYR; Biological variability in the structures of diphosphoinositol polyphosphates in Dictyostelium discoideum and mammalian cells. Biochem J 15 October 1997; 327 (2): 553–560. doi: https://doi.org/10.1042/bj3270553
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