Synthesis and characterization of non-hydrolysable diphosphoinositol polyphosphate messengers

Wu, Mingxuan; Dul, Barbara E.; Trevisan, Alexandra J.; Fiedler, Dorothea

doi:10.1039/c2sc21553e

Cited by 72 publications

(95 citation statements)

References 43 publications

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“…IP6K1-generated 5-IP7 modulates cell metabolism, signaling, exocytosis, and migration (48,(55)(56)(57)(58). IP6K1-generated 5-IP7 regulates insulin signaling by inhibiting the insulin-sensitizing protein kinase Akt both in vivo and in vitro (58)(59)(60)(61), whereas it promotes insulin secretion from pancreatic β cells (57). IP6K1 also regulates global transcription by 5-IP7-mediated inhibition of the histone demethylase JMJD2C (62).…”

Section: Resultsmentioning

confidence: 99%

Adipocyte-specific deletion of Ip6k1 reduces diet-induced obesity by enhancing AMPK-mediated thermogenesis

Zhu

Ghoshal

Rodrigues

et al. 2016

Journal of Clinical Investigation

156

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Section: Resultsmentioning

confidence: 99%

Adipocyte-specific deletion of Ip6k1 reduces diet-induced obesity by enhancing AMPK-mediated thermogenesis

Zhu

Ghoshal

Rodrigues

et al. 2016

Journal of Clinical Investigation

156

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“…To confirm that we were detecting cleavage of the pyrophosphate of 5PP-IP 5 , we tested the ability of Siw14 to utilize analogs of IP 7 that contain non-hydrolyzable methylene-bisphosphonate moieties (13,14). In these analogs, carbon replaces the oxygen in the anhydride linkage to the ␤-phosphate.…”

Section: Resultsmentioning

confidence: 99%

“…IP 6 was purchased from Sigma. Four isoforms of IP 7 (5-diphosphoinositol pentakisphosphate (5PP-IP 5 ), non-hydrolyzable 5-methylene-bisphosphonate inositol pentakisphosphate (5PCP-IP 5 ), 1PP-IP 5 , and non-hydrolyzable 1-methylene-bisphosphonate inositol pentakisphosphate (1PCP-IP 5 )) and IP 8 (3,5-bisdiphosphoinositol tetrakisphosphate (3,5PP-IP 4 )) were synthesized following the methods described previously (13)(14)(15).…”

Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

A Novel Inositol Pyrophosphate Phosphatase in Saccharomyces cerevisiae

Steidle

Chong

et al. 2016

Journal of Biological Chemistry

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Inositol pyrophosphates are high energy signaling molecules involved in cellular processes, such as energetic metabolism, telomere maintenance, stress responses, and vesicle trafficking, and can mediate protein phosphorylation. Although the inositol kinases underlying inositol pyrophosphate biosynthesis are well characterized, the phosphatases that selectively regulate their cellular pools are not fully described. The diphosphoinositol phosphate phosphohydrolase enzymes of the Nudix protein family have been demonstrated to dephosphorylate inositol pyrophosphates; however, the Saccharomyces cerevisiae homolog Ddp1 prefers inorganic polyphosphate over inositol pyrophosphates. We identified a novel phosphatase of the recently discovered atypical dual specificity phosphatase family as a physiological inositol pyrophosphate phosphatase. Purified recombinant Siw14 hydrolyzes the ␤-phosphate from 5-diphosphoinositol pentakisphosphate (5PP-IP 5 or IP 7 ) in vitro. In vivo, siw14⌬ yeast mutants possess increased IP 7 levels, whereas heterologous SIW14 overexpression eliminates IP 7 from cells. IP 7 levels increased proportionately when siw14⌬ was combined with ddp1⌬ or vip1⌬, indicating independent activity by the enzymes encoded by these genes. We conclude that Siw14 is a physiological phosphatase that modulates inositol pyrophosphate metabolism by dephosphorylating the IP 7 isoform 5PP-IP 5 to IP 6 .Inositol pyrophosphates are a novel class of signaling molecules that carry energy-rich diphosphate bonds. In wild-type yeast, the most abundant isoform, diphosphoinositol pentakisphosphate (PP-IP 5 or IP 7 ), 5 consists of a fully phosphorylated six carbon myo-inositol ring further pyrophosphorylated at one of the carbons. Two physiologically relevant isomers of IP 7 include 1PP-IP 5 and 5PP-IP 5 in which the pyrophosphate group is found at the 1-position or the 5-position, respectively. Sequential phosphorylation of IP 7 results in bisdiphosphoinositol tetrakisphosphate (1,5PP-IP 4 or IP 8 ) that is pyrophosphorylated at both the 1st and 5th positions (1, 2). Since their discovery more than 20 years ago, inositol pyrophosphates have been implicated in an array of processes, including cellular energetic metabolism, telomere maintenance, oxidative stress response, and vesicle trafficking in animals and fungi (1). Recent work in Saccharomyces cerevisiae found that production of inositol pyrophosphates is essential for mounting environmental stress responses (3). Inositol pyrophosphates are thought to be metabolic regulators displaying expanded cellular roles beyond classic second messengers (1).The enzymes that regulate the pools of inositol pyrophosphates are not fully described. The inositol hexakisphosphate kinases synthesize IP 7 , the most abundant inositol pyrophosphate in the cell; they add the ␤-phosphate to IP 6 at the 5th position generating 5PP-IP 5 (2). Yeast possess a single inositol hexakisphosphate kinase gene, KCS1, whereas mammals possess three homologous genes encoding inositol hexakisphosphate kinase (Fig....

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“…40 Although one report showed that IP 7 is not able to displace PI(3,4,5)P 3 from AKT in an in vitro binding assay, 77 it was later demonstrated that non-hydrolysable analogues of 5-IP 7 or 1-IP 7 can inhibit AKT activation even in the presence of PI(3,4,5)P 3 . 78 A recent study revealed that chemically synthesized 5-IP 7 released into mammalian cells is able to induce translocation of AKT from the plasma membrane to the cytoplasm. 79 It has been shown that AKT and other PH domains bind 5-IP 7 and IP 6 with higher affinity as compared with 1-IP 7 or IP 8 .…”

Section: Plasma Membranementioning

confidence: 99%

“…78,79,[126][127][128][129] Highly pure PP-IPs with β-phosphate moieties at specific carbon atoms have been synthesized by these groups, including non-hydrolysable analogues that are stable in cells and can bind target proteins but not pyrophosphorylate them. 47,78,130 A system for intracellular delivery and photouncaging of chemically synthesized PP-IPs has recently been developed, 79 which promises to open up new methods to study the functions of these molecules in different subcellular compartments. The use of IP 7 as an affinity reagent revealed two different classes of interacting proteins, depending on the absence or presence of the divalent cation Mg 2+ during the interaction, representing IP 7 binding or pyrophosphorylation targets respectively.…”

Section: Perspective On the Futurementioning

confidence: 99%

Inositol Pyrophosphates: Energetic, Omnipresent and Versatile Signalling Molecules

et al. 2017

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| Inositol pyrophosphates (PP-IPs) are a class of energy-rich signalling molecules found in all eukaryotic cells. These are derivatives of inositol that contain one or more diphosphate (or pyrophosphate) groups in addition to monophosphates. The more abundant and best studied PP-IPs are diphosphoinositol pentakisphosphate (IP 7 ) and bis-diphosphoinositol tetrakisphosphate (IP 8 ). These molecules can influence protein function by two mechanisms: binding and pyrophosphorylation. The former involves the specific interaction of a particular inositol pyrophosphate with a binding site on a protein, while the latter is a unique attribute of inositol pyrophosphates, wherein the β-phosphate moiety is transferred from a PP-IP to a pre-phosphorylated serine residue in a protein to generate pyrophosphoserine. Both these events can result in changes in the target protein's activity, localisation or its interaction with other partners. As a consequence of their ubiquitous presence in all eukaryotic organisms and all cell types examined till date, and their ability to modify protein function, PP-IPs have been found to participate in a wide range of metabolic, developmental, and signalling pathways.This review highlights many of the known functions of PP-IPs in the context of their temporal and spatial distribution in eukaryotic cells. The pathway of synthesis of inositol polyphosphates has been characterized in yeast, slime moulds, plants and animals. The simplest anabolic pathway, characterized in the yeast Saccharomyces cerevisiae (Fig. 1) (Fig. 1). Interestingly, the PPIP5Ks also possess a phosphatase domain which selectively cleaves the 1-position β-phosphate of 1-IP 7 and IP 8 , thus targeting the products of the kinase domain.35, 36 A recent study identified another yeast phosphatase, Siw14, that specifically targets the 5-position β-phosphate of PP-IPs 37 (Fig. 1). As PP-IPs are found in all eukaryotic organisms, they display many conserved and divergent 2, 131 Siw14, an inositol pyrophosphate phosphatase in yeast, preferentially cleaves the C5 β-phosphate on PP-IPs. 37 The yeast enzymes are depicted in purple, and mammalian enzymes are depicted in green and are bracketed. The undetermined inositol pyrophosphate structure is represented with an interrogation mark. Myoinositol contains five equatorial (parallel to the axis) and one axial (perpendicular to the axis) hydroxyl groups. Carbon atoms on the myo-inositol ring are numbered on the structures of PI(4,5)P 2 and IP 6 .

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Synthesis and characterization of non-hydrolysable diphosphoinositol polyphosphate messengers

Cited by 72 publications

References 43 publications

Adipocyte-specific deletion of Ip6k1 reduces diet-induced obesity by enhancing AMPK-mediated thermogenesis

Adipocyte-specific deletion of Ip6k1 reduces diet-induced obesity by enhancing AMPK-mediated thermogenesis

A Novel Inositol Pyrophosphate Phosphatase in Saccharomyces cerevisiae

Inositol Pyrophosphates: Energetic, Omnipresent and Versatile Signalling Molecules

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