Structural analysis and detection of biological inositol pyrophosphates reveals that the VIP/PPIP5K family are 1/3-kinases.

TY - JOUR

T1 - Structural analysis and detection of biological inositol pyrophosphates reveals that the VIP/PPIP5K family are 1/3-kinases.

AU - Lin, Hongying

AU - Fridy, Peter C

AU - Ribeiro, Anthony A

AU - Choi, Jae H

AU - Barma, Deb K

AU - Vogel, Günter

AU - Falck, J R

AU - Shears, Stephen B

AU - York, John D

AU - Mayr, Georg W.

PY - 2009

Y1 - 2009

N2 - We have characterized the positional specificity of the mammalian and yeast VIP/PPIP5K family of inositol phosphate kinases. We deployed a micro-scale 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 phosphorylate 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-diphosphoinositol pentakisphosphate. The latter compound has not previously been identified in any organism. We have also unequivocally determined that 1/3,5-(PP)2-IP4 is the isomeric structure of the bis-diphosphoinositol tetrakisphosphate that is synthesized by yeasts and mammals, through a collaboration between the IP6K 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-IP4 (Laussmann et al., Biochem. J. 1996 315 715-720). 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.

AB - We have characterized the positional specificity of the mammalian and yeast VIP/PPIP5K family of inositol phosphate kinases. We deployed a micro-scale 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 phosphorylate 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-diphosphoinositol pentakisphosphate. The latter compound has not previously been identified in any organism. We have also unequivocally determined that 1/3,5-(PP)2-IP4 is the isomeric structure of the bis-diphosphoinositol tetrakisphosphate that is synthesized by yeasts and mammals, through a collaboration between the IP6K 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-IP4 (Laussmann et al., Biochem. J. 1996 315 715-720). 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.

M3 - SCORING: Zeitschriftenaufsatz

VL - 284

SP - 1863

EP - 1872

JO - J BIOL CHEM

JF - J BIOL CHEM

SN - 0021-9258

IS - 3

M1 - 3

ER -