Structural analysis and detection of biological inositol pyrophosphates reveal that the family of VIP/Diphosphoinositol pentakisphosphate kinases Are1/3-kinases

Hongying Lin, Peter C. Fridy, Anthony A. Ribeiro, Jae H. Choi, Deb K. Barma, Günter Vogel, J. R. Falck, Stephen B. Shears, John D. York, Georg W. Mayr

Research output: Contribution to journalArticle

83 Scopus citations

Abstract

We have characterized the positional specificity of the mammalian and yeast VIP/diphosphoinositol pentakisphos-phate kinase (PPIP5K) family of inositol phosphate kinases. We deployed a microscale metal dye detection protocol coupled to a high performance liquid chromatography system that was calibrated with synthetic and biologically synthesized standards of inositol pyrophosphates. In addition, we have directly analyzed the structures of biological inositol pyrophosphates using two-dimensional 1H- 1H and 1H- 31P nuclear magnetic resonance spectroscopy. Using these tools, we have determined that the mammalian and yeast VIP/PPIP5K family phosphorylates the 1/3-position of the inositol ring in vitro and in vivo. For example, the VIP/PPIP5K enzymes convert inositol hexakisphosphate to 1/3-diphos-phoinositol pentakisphosphate. The latter compound has not previously been identified in any organism. We have also unequivocally determined that 1/3,5-(PP) 2-IP 4 is the isomeric structure of the bis-diphosphoinositol tetrakisphos-phate that is synthesized by yeasts and mammals, through a collaboration between the inositol hexakisphosphate kinase and VIP/PPIP5K enzymes. These data uncover phylogenetic variability within the crown taxa in the structures of inositol pyrophosphates. For example, in the Dictyostelids, the major bis-diphosphoinositol tetrakisphosphate is 5,6-(PP) 2-IP 4 (Laussmann, T., Eujen, R., Weisshuhn, C. M., Thiel, U., Falck, J. R., and Vogel, G. (1996) Biochem. J. 315, 715-725). Our study brings us closer to the goal of understanding the structure/function relationships that control specificity in the synthesis and biological actions of inositol pyrophosphates.

Original languageEnglish (US)
Pages (from-to)1863-1872
Number of pages10
JournalJournal of Biological Chemistry
Volume284
Issue number3
DOIs
StatePublished - Jan 16 2009

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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