SHEARS SB. Inositolipyrofosfaatit; (InsPP) Miksi niin monta fosfaattia inositolijärjestelmällä?

https://www.ncbi.nlm.nih.gov/pubmed/25453220

Adv Biol Regul. 2015 Jan;57:203-16. doi: 10.1016/j.jbior.2014.09.015. Epub 2014 Oct 5.

Inositol pyrophosphates: why so many phosphates?

Shears SB¹.

1

Inositol Signaling Group, Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, NIH, DHHS, PO Box 12233, Research Triangle Park, NC 27709, USA. Electronic address: Shears@niehs.nih.gov.

Abstract

The inositol pyrophosphates (PP-InsPs) are a specialized group of "energetic" signaling molecules found in yeasts, plants and animals. PP-InsPs boast the most crowded three dimensional phosphate arrays found in Nature; multiple phosphates and diphosphates are crammed around the six-carbon, inositol ring. Yet, phosphate esters are also a major energy currency in cells. So the synthesis of PP-InsPs, and the maintenance of their levels in the face of a high rate of ongoing turnover, all requires significant bioenergetic input. What are the particular properties of PP-InsPs that repay this investment of cellular energy? Potential answers to that question are discussed here, against the backdrop of a recent hypothesis that signaling by PP-InsPs is evolutionarily ancient. The latter idea is extended herein, with the proposal that the primordial origins of PP-InsPs is reflected in the apparent lack of isomeric specificity of certain of their actions. Nevertheless, there are other aspects of signaling by these polyphosphates that are more selective for a particular PP-InsP isomer. Consideration of the nature of both specific and non-specific effects of PP-InsPs can help rationalize why such molecules possess so many phosphates.

Published by Elsevier Ltd.

KEYWORDS:

Analogs; Cell-signaling; Diphosphoinositol polyphosphates; Inositol pyrophosphates; Kinase; Phosphorylation; Structure

PMID:

25453220

PMCID:

PMC4291286

DOI:

10.1016/j.jbior.2014.09.015

[Indexed for MEDLINE]

Free PMC Article

KOSKA ON INOSITOLIPYROFOSFAATTEJA, on myös INOSITOLIPYROFOSFATAASEJA:

TAKESHI IJUIN (2019) artikkeli ja kuvat inositoliaineenvaihdunnan fosfataaseista mm. Syöpäsoludynamiikkaa pohditaan.

 Koska inositoliaineenvaihdunta  kiertää   vesiliukoisesta  ja rasvaliukoiseen aitioon  ja takaisin  on  fosfataaseissa niitä, jotka  vaikuttavat a)   lipidiliukoisiin fosfoinositideihin (PI, lipositolit) ja  b)  vesiliukoisiin  jonistolifosfaattilajeihin (IPx)
 Sisällysluettelossa on  seuraavia vesiliukoisia fosfataaseja mainittu:
INPP4A, Inositolipolyfosfaatti 4-fosfataasi tyyppi 1
INPP4B, Inositolipolyfosfaatti 4-fosfataasi tyyppi 2
INPP5J, Inositolipolyfosfaatti 5-fosfataasi J
INPP5K, Inositolipolyfosfatti 5-fosfataasi K
PIPP, proliinipitoinen inositolipolyfosfaattifosfataasi 

SHIP1,  SH2 domeenin sisältävä  inositolifosfaasi 1

SHIP2,  SH2 domeenin  sisältävä isositolifosfataasi  2

SKIP, lihakseen ja munuaiseen rikastunut inositol polyfosfaattifosfataasi

Rasvaliukoiseen  fosfoinositidiin vaikuttaa
PTEN , fosfataasi ja tensiinihomologi
PTPN9, PTP-MEG2 (T-soluissa polyfosfoinositideihin esim.)
Myotubulariini MTMR2 on fosfoinositidifosfataasi .

https://www.sciencedirect.com/science/article/pii/S1044579X18301159?via%3Dihub

Review

Phosphoinositide phosphatases in cancer cell dynamics—Beyond PI3K and PTEN

Author links open overlay panel TakeshiIjuin

https://doi.org/10.1016/j.semcancer.2019.03.003Get rights and content

Under a Creative Commons license

open access

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Abstract

Phosphoinositides (PIs)  are a group of lipids that regulate intracellular signaling and subcellular biological events. The signaling by PIP3,  phosphatidylinositol-3,4,5-trisphosphate and Akt mediates the action of growth factors that are essential for cell proliferation, gene transcription, cell migration, and polarity. The hyperactivation of this signaling has been identified in different cancer cells; and, it has been implicated in oncogenic transformation and cancer cell malignancy. Recent studies have argued the role of phosphoinositides  (PIs) in cancer cell dynamics, including actin cytoskeletal rearrangement at the plasma membrane and the organization of intracellular compartments. The focus of this review is to summarize the impact of the activities of phosphoinositide (PI) phosphatases on intracellular signaling related to cancer cell dynamics and to discuss how the abnormalities in the activities of the enzymes alter the levels of phosphoinositides (PIs, lipositols) in cancer cells.

Abbreviations

ER.   endoplasmic reticulum

ERM.   ezrin, radixin, and Moesin

ESCC.  esophageal squamous cell carcinoma

FA. focal adhesion

FAK. focal adhesion kinase

FEME. fast endophilin-mediated endocytosis

GOLPH3. Golgi phosphoprotein 3

INPP4A. inositol polyphosphate 4-phosphatase type I

INPP4B. inositol polyphosphate 4-phosphatase type II

Lpd. lamellipodin

INPP5J. inositol polyphosphate 5-phosphatase J

INPP5K. inositol polyphosphate 5-phosphatase K

LOH. loss of heterogeneity

N-WASP. neural Wiskott–Aldrich syndrome protein

PDK1. phosphoinositide-dependent kinase 1

PH. plextrin homology

PI. phosphatidylinositol

PI(3)P. phosphatidylinositol-3-monophosphate

PI(4)P. phosphatidylinositol-4-monophosphate

PI(5)P. phosphatidylinositol-5-monophosphate

PI(3,4)P_2. phosphatidylinositol-3,4-bisphosphate

PI(3,5)P_2. phosphatidylinositol-3,5-bisphosphate

PI(4,5)P_2. phosphatidylinositol-4,5-bisphosphate

PIP_3. phosphatidylinositol-3,4,5-trisphosphate

PI3K. PI 3-kinase

PI4K. phosphatidylinositol 4-kinase

PIP5K. phosphatidylinositol 4-phosphate 5-kinase

PIPP. proline-rich inositol polyphosphate 5-phosphatase

PM. plasma membrane

P-Rex. phosphatidylinositol 3,4,5-trisphosphate-dependent Rac exchanger

PTEN. phosphatase and tensin homolog

PX. phox homology

SAC. suppressor of actin

SHIP1,  SH2 domain-containing inositol phosphatase 1

SHIP2,  SH2 domain-containing inositol phosphatase 2

SKIP,  skeletal muscle and kidney enriched inositol polyphosphate phosphatase

SYNJ2,  synaptojanin 2

Tks5,  tyrosine kinase substrate 5

Keywords

Cell migration

Cell polarity

Focal adhesion

Invadopodia

PI(3,4)P₂

PIP₃

Phosphoinositide phosphatase

CHEN MJ (2017) .Proteiinifosfataasien genomi ja evoluutio.

https://www.ncbi.nlm.nih.gov/pubmed/28400531

Sci Signal. 2017 Apr 11;10(474). pii: eaag1796. doi: 10.1126/scisignal.aag1796.

Genomics and evolution of protein phosphatases.

Chen MJ^1,2, Dixon JE³, Manning G^4,2.

1

Department of Bioinformatics and Computational Biology, Genentech Inc., South San Francisco, CA 94080, USA.

2

Razavi Newman Center for Bioinformatics, Salk Institute for Biological Studies, La Jolla, CA 94027, USA.

3

Departments of Pharmacology, Cellular and Molecular Medicine, and Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093, USA.

4

Department of Bioinformatics and Computational Biology, Genentech Inc., South San Francisco, CA 94080, USA. manning@manninglab.org.

Tiivistelmä  Abstract

 PROTEIINIFOSFATAASIT ovat essentiellejä proteiinikinaasien opponentteja.
Yhdessä nämä entsyymit säätelevät kaikkien proteiinien fosforyloitumisen ja useimmat soluprosessit.  Jota voidaan paremmin käsittää proteiinifosforylaation  yleisosuutta, tutkijat  tekivät luettelon ihmisproteiinioen fosfatomista, joka käsittää 189  tunnettua ja oletettua ihmisen proteiinifosfataasigeeniä.  He tunnistivat myös 79 proteiinifosfataasipseudogeeniä tai retrogeeniä, joista joillakin säättaa olla jäännösfunktioita. He jäöjittivät fosfataasien  alkuperää ja diversiteettiä  rakentamalla proteiinifosfatomeja kahdeksasta muusta  eukaryosyytistä  protistista Dictyostelium  merisiiliin ( sea urchin) .  He luokittelivat  proteiinifosfataasit  kaikista yhdeksästä lajista kymmenen proteiinilaskoksen, 21 perheen ja 178 alaperheen hierarkiaan. he havaitsivat, että  yli 80% ihmisalaperheistä oli konservoituneita kautta  eläinkunnan. , mutta osoitti huomattavaaeroavuuksia  evoluutiossa, kuten ksittäisten alaperheiden  katoamisia ja  laajenemisia ja muutoksia liitedomeeneissa.  Proteiinifosfataasit osoittavat  samankaltaista  evolutionaalista dynamiikkaa kuin  proteiinikinaasit, huomattavin   katoamisin useimmissa  malliorganismeissa. Sekvenssianalyysin mukaan voidaan olettaa, että 26 ihmsen  proteiinifosfataasidomeenia on   menettänyt katalyyttisen kyvyn ja tälläionen  kyvyttömyys  on konservoituneinta  kautta ortologien.  Tällainen genominen ja evolutionaarinen näkökohta proteiinifosfataasesita antaa  kehiötä  eläinkunnan  proteiinifosforylaation yleisanalyysiin.

Protein phosphatases are the essential opposite to protein kinases; together, these enzymes regulate all protein phosphorylation and most cellular processes. To better understand the global roles of protein phosphorylation, we cataloged the human protein phosphatome, composed of 189 known and predicted human protein phosphatase genes. We also identified 79 protein phosphatase pseudogenes or retrogenes, some of which may have residual function. We traced the origin and diversity of phosphatases by building protein phosphatomes for eight other eukaryotes, from the protist Dictyostelium to the sea urchin. We classified protein phosphatases from all nine species into a hierarchy of 10 protein folds, 21 families, and 178 subfamilies. We found that >80% of the 101 human subfamilies were conserved across the animal kingdom, but show substantial differences in evolution, including losses and expansions of individual subfamilies and changes in accessory domains. Protein phosphatases show similar evolutionary dynamics to those of kinases, with substantial losses in major model organisms. Sequence analysis predicts that 26 human protein phosphatase domains are catalytically disabled and that this disability is mostly conserved across orthologs. This genomic and evolutionary perspective on protein phosphatases provides a framework for global analysis of protein phosphorylation throughout the animal kingdom.

Copyright © 2017, American Association for the Advancement of Science.

PMID:

28400531

DOI:

10.1126/scisignal.aag1796

[Indexed for MEDLINE]

CARMAN GM (1997). DAG- pyrofosfataasi, PA fosfataasi(fosfatidihappofosfataasi) .

https://www.ncbi.nlm.nih.gov/pubmed/9370315

Phosphatidate phosphatases and diacylglycerol pyrophosphate phosphatases in Saccharomyces cerevisiae and Escherichia coli.

Carman GM¹.

1

Department of Food Science, Cook College, New Jersey Agricultural Experiment Station, Rutgers University, New Brunswick 08903, USA. carman@aesop.rutgers.edu

Abstract

Phosphatidate phosphatase  (PA phosphatase) plays a major role in the synthesis of phospholipids and triacylglycerols in the yeast Saccharomyces cerevisiae. Membrane- and cytosolic-associated forms of the enzyme have been isolated and characterized. These enzymes are Mg2+-dependent and N-ethylmaleimide-sensitive. The expression of a membrane-associated form of phosphatidate phosphatase is regulated by growth phase and inositol supplementation, whereas enzyme activity is regulated by lipids, nucleotides, and by phosphorylation. Phosphatidate phosphatase is coordinately regulated with other phospholipid biosynthetic enzymes including phosphatidylserine synthase.

Diacylglycerol pyrophosphate phosphatase   is a novel enzyme of phospholipid metabolism which is present in S. cerevisiae, Escherichia coli, and mammalian cells. This enzyme possesses a phosphatidate phosphatase activity which is Mg2+-independent and N-ethylmaleimide-insensitive and is distinct from the Mg2+-dependent and N-ethylmaleimide-sensitive form of phosphatidate phosphatase.

 Genes encoding for diacylglycerol pyrophosphate phosphatase have been isolated from S. cerevisiae and E. coli. The deduced protein sequences of these genes show homology to the sequence of the mouse PAP2 (Mg2+-independent and N-ethylmaleimide-insensitive phosphatidate phosphatase) protein, especially in a novel phosphatase sequence motif. Rat liver PAP2 displays diacylglycerol pyrophosphate phosphatase activity.

PMID:

9370315

DOI:

10.1016/s0005-2760(97)00095-7

[Indexed for MEDLINE]

DAG PPi fosfataasit:

PLPP7,

https://www.ncbi.nlm.nih.gov/gene/?term=Homo+sapiens+PLPP7

Official Symbol

PLPP7provided by HGNC

Official Full Name

phospholipid phosphatase 7 (inactive)provided by HGNC

Also known as

NET39; C9orf67; PPAPDC3

Expression

Biased expression in heart (RPKM 14.9), brain (RPKM 2.9) and 9 other tissues See more

Orthologs

mouse all

Preferred Names

inactive phospholipid phosphatase 7

Names

epididymis secretory sperm binding protein

nuclear envelope transmembrane protein NET39

phosphatidic acid phosphatase type 2 domain containing 3

phosphatidic acid phosphatase type 2 domain-containing protein 3

probable lipid phosphate phosphatase PPAPDC3

 https://www.ncbi.nlm.nih.gov/pubmed/19704009/

Muscle homeostasis  

PLPP6,

  https://www.ncbi.nlm.nih.gov/gene/403313

Official Full Name

phospholipid phosphatase 6provided by HGNC

Also known as

PDP1; PSDP; PPAPDC2; bA6J24.6

Preferred Names

phospholipid phosphatase 6

Names

PPAP2 domain-containing protein 2

phosphatidic acid phosphatase type 2 domain containing 2

phosphatidic acid phosphatase type 2 domain-containing protein 2

polyisoprenoid diphosphate phosphatase type 1

presqualene diphosphate phosphatase (PSDP)

findings indicate that PPAPDC2 in human PMN is the first lipid phosphate phosphohydrolase identified for PSDP; regulation of this activity of the enzyme may have important roles for PMN activation in innate immunity

PLPP5 

Official Symbol

PLPP5provided by HGNC

Official Full Name

phospholipid phosphatase 5 provided by HGNC

https://www.ncbi.nlm.nih.gov/gene/?term=(DPPL1%3B+HTPAP%3B+PPAPDC1BM
DPPL1; HTPAP; PPAPDC1BM

Preferred Names

phospholipid phosphatase 5

Names

diacylglycerol pyrophosphate like 1

diacylglycerol pyrophosphate phosphatase-like 1

phosphatidate phosphatase PPAPDC1B

phosphatidic acid phosphatase type 2 domain containing 1B

phosphatidic acid phosphatase type 2 domain-containing protein 1B

testicular secretory protein Li 38

 PLPP4    ( expression BRAIN)

Official Symbol

PLPP4provided by HGNC

Official Full Name

phospholipid phosphatase 4provided by HGNC

Primary source

HGNC:HGNC:23531

See related

Ensembl:ENSG00000203805

Gene type

protein coding

RefSeq status

VALIDATED

Organism

Homo sapiens

Lineage

Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini; Catarrhini; Hominidae; Homo

Also known as

DPPL2; PPAPDC1; PPAPDC1A

Expression

Low expression observed in reference dataset See more

Orthologs

mouse all

Preferred Names

phospholipid phosphatase 4

Names

diacylglycerol pyrophosphate like 2

diacylglycerol pyrophosphate phosphatase-like 2

phosphatidate phosphatase PPAPDC1A

phosphatidic acid phosphatase type 2 domain containing 1A

phosphatidic acid phosphatase type 2 domain-containing protein 1A

 PLPP3 , Phospholipid phosphatase 3

https://www.ncbi.nlm.nih.gov/gene/8613

Also known as

LPP3; VCIP; Dri42; PAP2B; PPAP2B

Summary

The protein encoded by this gene is a member of the phosphatidic acid phosphatase (PAP) family. PAPs convert phosphatidic acid to diacylglycerol, and function in de novo synthesis of glycerolipids as well as in receptor-activated signal transduction mediated by phospholipase D. This protein is a membrane glycoprotein localized at the cell plasma membrane. It has been shown to actively hydrolyze extracellular lysophosphatidic acid and short-chain phosphatidic acid. The expression of this gene is found to be enhanced by epidermal growth factor in Hela cells. [provided by RefSeq, Mar 2010]

Expression

Broad expression in thyroid (RPKM 205.6), placenta (RPKM 102.2) and 23 other tissues See more

Preferred Names

phospholipid phosphatase 3

Names

PAP2 beta

lipid phosphate phosphohydrolase 3

phosphatidate phosphohydrolase type 2b

phosphatidic acid phosphatase type 2B

type-2 phosphatidic acid phosphatase-beta

vascular endothelial growth factor and type I collagen inducible

We identified vascular endothelial growth factor and type I collagen inducible protein (VCIP), also known as phosphatidic acid phosphatase 2b (PAP2b), in a functional assay of angiogenesis. 

  PLPP2

 https://www.ncbi.nlm.nih.gov/gene/8612

Also known as

LPP2; PAP-2c; PAP2-g; PPAP2C

Preferred Names

phospholipid phosphatase 2

Names

PAP2-gamma

PAP2c

lipid phosphate phosphohydrolase 2, (LPP2)

phosphatidate phosphohydrolase type 2c

phosphatidic acid phosphatase 2c

phosphatidic acid phosphatase type 2C, (PPAP2C)

phosphatidic acid phosphohydrolase type 2c

type-2 phosphatidic acid phosphatase-gamma,  (PAP2-g)

The lung contains two distinct forms of phosphatidic acid phosphatase (PAP). PAP1 is a cytosolic enzyme that is activated through fatty acid-induced translocation to the endoplasmic reticulum, where it converts phosphatidic acid (PA) to diacylglycerol (DAG) for the biosynthesis of phospholipids and neutral lipids. PAP1 is Mg(2+) dependent and sulfhydryl reagent sensitive. PAP2 is a six-transmembrane-domain integral protein localized to the plasma membrane. Because PAP2 degrades sphingosine-1-phosphate (S1P) and ceramide-1-phosphate in addition to PA and lyso-PA, it has been renamed lipid phosphate phosphohydrolase (LPP). LPP is Mg(2+) independent and sulfhydryl reagent insensitive. This review describes LPP isoforms found in the lung and their location in signaling platforms (rafts/caveolae). Pulmonary LPPs likely function in the phospholipase D pathway, thereby controlling surfactant secretion. Through lowering the levels of lyso-PA and S1P, which serve as agonists for endothelial differentiation gene receptors, LPPs regulate cell division, differentiation, apoptosis, and mobility. LPP activity could also influence transdifferentiation of alveolar type II to type I cells. It is considered likely that these lipid phosphohydrolases have critical roles in lung morphogenesis and in acute lung injury and repair.

PLPP1

https://www.ncbi.nlm.nih.gov/gene/8611 

Official Symbol

PLPP1provided by HGNC

Official Full Name

phospholipid phosphatase 1provided by HGNC

Primary source

HGNC:HGNC:9228

See related

Ensembl:ENSG00000067113 MIM:607124

Gene type

protein coding

RefSeq status

REVIEWED

Organism

Homo sapiens

Lineage

Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini; Catarrhini; Hominidae; Homo

Also known as

LPP1; PAP2; LLP1a; PAP-2a; PPAP2A

Summary

The protein encoded by this gene is a member of the phosphatidic acid phosphatase (PAP) family. PAPs convert phosphatidic acid (PA)  to diacylglycerol (DAG), and function in synthesis of glycerolipids and in phospholipase D-mediated signal transduction. This enzyme is an integral membrane glycoprotein that plays a role in the hydrolysis and uptake of lipids from extracellular space. Alternate splicing results in multiple transcript variants of this gene. [provided by RefSeq, May 2013]

Expression

Broad expression in prostate (RPKM 96.2), gall bladder (RPKM 74.3) and 22 other tissues See more

Orthologs

mouse all

Preferred Names

phospholipid phosphatase 1

Names

lipid phosphate phosphohydrolase 1a

phosphatidate phosphohydrolase type 2a

phosphatidic acid phosphatase 2a

phosphatidic acid phosphatase type 2A

phosphatidic acid phosphohydrolase type 2a

type-2 phosphatidic acid phosphatase alpha

ZHAO H (2019).Pyrofosfataasien havaitseminen fluoresoivalla menetelmllä.

https://www.ncbi.nlm.nih.gov/pubmed/30572018

Mol Cell Probes. 2019 Feb;43:29-33. doi: 10.1016/j.mcp.2018.12.003. Epub 2018 Dec 17.

A novel fluorometric method for inorganic pyrophosphatase detection based on G-quadruplex-thioflavin T.

Zhao H¹, Ma C², Chen M¹.

1

School of Life Sciences, Central South University, Changsha, 410013, China.

2

School of Life Sciences, Central South University, Changsha, 410013, China. Electronic address: macb2012@csu.edu.cn.

Abstract

In this paper, we propose a fluorometric approach for the highly sensitive detection of inorganic pyrophosphatase (PPase) based on G-quadruplex-thioflavin T (ThT). In the absence of PPase, Cu²⁺ can coordinate with pyrophosphate (PPi) to generate a Cu²⁺/PPi complex. Then the G-rich sequence folds into the G-quadruplex structure, which can combine with ThT to generate a remarkable fluorescent signal. In the presence of PPase, the coordinated compound can be destroyed by the PPase catalyzed hydrolysis of PPi into inorganic phosphate (Pi). The subsequent release of Cu²⁺ can compete with ThT to induce a tighter G-quadruplex structure, causing the release of ThT and a sharp fluorescence decrease. Based on this mechanism, a facile and quantitative strategy for PPase detection was developed. The fluorescence intensity of the system shows a linear relationship with the PPase activities in the range of 0.5-30 U/L with a detection limit as low as 0.48 U/L. The proposed strategy for fluorescence spectrometric PPase detection is convenient, cost effective, and sensitive. This can be utilized to evaluate the inhibition effect of NaF on PPase as well as diagnose PPase-related diseases.

Copyright © 2018 Elsevier Ltd. All rights reserved.

KEYWORDS:

Fluorescence; G-quadruplex-thioflavin T; Label-free; Pyrophosphatase

PMID:

30572018

DOI:

10.1016/j.mcp.2018.12.003

[Indexed for MEDLINE]

WANG Y (2018) Kuinka voi havaita pyrofosfataaseja? Uusi elektrokemiallinen menetelmä luotu 2017.

yksi syy miksi fosfaattien tutkimus on erittäin aktuellia;:  havaitsemistekniikka tästä  fosforin ja  energia-alueella  signalointia suorittavien proteiinien ja  mineraalien   ja fosforiyhdisteiden dynamiikasta  on  suuresti kehittynyt  vuosituhannen vaihteen jälkeen.  Esim  fytiinin  aineenvaihdunt mudoosta  vähän väliä pyrofosfaatteja samoin kuin sokeriaineenvaihdunta.  Entsyymit, jotka sitten  katalysoivat  jonien "salamannopeaita " ajallisia ja  virrallisia  vektoreita,  ovat nyt vuorossa tulla havaituiksi.

https://www.ncbi.nlm.nih.gov/pubmed/29136853

• Format: Abstract

Talanta. 2018 Feb 1;178:491-497. doi: 10.1016/j.talanta.2017.09.069. Epub 2017 Sep 28.

Electrochemical strategy for pyrophosphatase detection Based on the peroxidase-like activity of G-quadruplex-Cu²⁺ DNAzyme.

Wang Y¹, Wu Y², Liu W³, Chu L³, Liao Z³, Guo W³, Liu GQ⁴, He X⁵, Wang K⁶.

1

Hunan Provincial Key Laboratory for Forestry Biotechnology, College of Life Science and Technology, Central South University of Forestry and Technology, 410004 Changsha, PR China; State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biology, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: bionano@163.com.

2

Hunan Provincial Key Laboratory for Forestry Biotechnology, College of Life Science and Technology, Central South University of Forestry and Technology, 410004 Changsha, PR China; State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biology, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.

3

Hunan Provincial Key Laboratory for Forestry Biotechnology, College of Life Science and Technology, Central South University of Forestry and Technology, 410004 Changsha, PR China.

4

Hunan Provincial Key Laboratory for Forestry Biotechnology, College of Life Science and Technology, Central South University of Forestry and Technology, 410004 Changsha, PR China. Electronic address: gaoliuedu@163.com.

5

State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biology, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.

6

State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biology, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: kmwang@hnu.edu.cn.

 TIIVISTELMÄ, Abstract

 Uusi yksinkertainen ja hyvin herkkä elektrokemiallinen metodi pyrofosfataasiaktiivisuuden havaitsemiseen  on kehitetty  ja se perustuu G-quadruplex-Cu2+ DNAzyymin peroksidaasinkaltaiseen aktiivisuuteen.
Kun  pyrofosfataasientsyymiä ei ole läsnä, Cu2+ voisi koordinoitua pyrofosfaatin (PPi) kanssa  muodostaen  Cu2+-PPi yhdistyksen. Mutta jos on läsnä  pyrofosfataasia (PPaasi),  entsyymi  voisi hävittää  koordinoituneen yhdisteen, koska  PPaasi katalysoi pyrofosfaatin (PPi) hydrolyysin epäorgaaniseksi fosfaatiksi Pi ja tuottaa vapaata  Cu2+ jonia, joka sitten pystyy kytkeytymään G-pitoiseen DNA:han muodostaen G-quadruplex-Cu2+DNAzyymin.   G-quadruplex-Cu2+DNAzyymin muodostuminen  immobilisoitui SPGE-pinnalla (kultaelektrodilla).  Kun käytettiin   redox-väliaineena TMB ja vetyperoksidia (H2O2 entsyymisubstraattina ,DNAzyymi katalysoi vetyperoksidin reduktion kehkeyttäen mitattavan,  kvantitatiivisen kronoamperometrisen signaalin.  Mainitun G-quadruplex-Cu2+-DNAzyymin  katalyyttinen aktiviteetti TMB-H2O2- reaktiossa oli suhteessa  pyrofosfataasin aktiivisuuteen - ja täten oli saatu  ensimmäistä kertaa  kehitettyä  yksinkertainen  ja herkkä  pyrofosfataasiaktiivisuudesta riippuva  elektrokemiallinen metodi. Järjestelmän kronoamperometrinen intensiteetti omaa eräänlaisen lineaarin suhteen  pyrofosfataasin aktiivisuuksiin rajoissa 1.0- 50.0 mU/mL.  ja havaitsemisen raja  voisi olla niinkin alhaalla kuin 0.6 mU/mL (S/N=3). Tämä esitetty metodi oli selektiivinen,  taloudellinen ja soveltuvainen,  omaa laatuaan,  ilman  monimutkaisia operaatioita   ja sitä on edelleen sovellettu  pyrofosfataasien inhibiitorien seulontaan hyvin tehokkaasti. 

A new simple and highly sensitive electrochemical method for pyrophosphatase (PPase) activity detection was developed based on the peroxidase-like activity of G-quadruplex-Cu²⁺ DNAzyme. In the absence of PPase, Cu²⁺ could coordinate with pyrophosphate (PPi) to form Cu²⁺-PPi compound. While in the presence of PPase, it could destroy the coordinate compound because PPase catalyzed the hydrolysis of PPi into inorganic phosphate and produced free Cu²⁺, which then could be coupled with G-rich DNA to form G-quadruplex-Cu²⁺ DNAzyme. The formation of a mimic enzyme (G-quadruplex-Cu²⁺ DNAzyme) was immobilized on the surface of screen-printed gold electrode (SPGE). Using 3, 3', 5, 5'-tetramethylbenzidine (TMB) as a redox mediator and H₂O₂ as an enzyme substrate, the DNAzyme catalyzed the reduction of H₂O₂ to generate quantitative chronoamperometric signal. The catalytic activity of G-quadruplex-Cu²⁺ DNAzyme for TMB-H₂O₂ reaction was proportional to the activity of PPase, based on which, a simple and sensitive turn-on electrochemical method for PPase activity was thus developed for the first time. The chronoamperometric intensity of the system had a linear relationship with the PPase activities in the range of 1.0-50.0mU/mL and the detection limit could be down to 0.6mU/mL (S/N = 3). This proposed method was selective, cost-effective and convenient without any labels or complicated operations, which was furthermore applied to screen the inhibitor for PPase with high efficiency.

Copyright © 2017. Published by Elsevier B.V.

KEYWORDS:

Chronoamperometric detection; DNAzyme; G-Quadruplex; Peroxidase-like activity; Pyrophosphatase

PMID:

29136853

DOI:

10.1016/j.talanta.2017.09.069

[Indexed for MEDLINE]

• 
  

Miten monta PTP fosfataasia osallistuu inositolifosfaattien aineenvaihduntaverkostoon?

 Miksi on niin paljon proteiinityrosiinifosfataaseja,PTP?

 Ihmisen genomissa on  proteiinityrosiinifosfataasigeenejä (PTP)  enemmän kuin proteiinityrosiinikinaasigeenejä (PTK). PTP geenejä on 107 ja  tyrosiinikinaasigeenejä 90. 

PTP-geeneistä on katalyyttisesti inaktiiveja 11 geenia. Kaksi geeniä defosforyloi lähetti-RNA:ta (PIR1 mRNA defosforylaasi; Geeni DUSP11; mRNA capping enzyme, geeni RNGTT)

 ja 13 PTP entsyymiä    toimii inositolifosfolipideihin  (PI)   rajoitetulla alueella  niitä defosforyloimassa ( Koetan löytää tämän joukon:  PTEN geeniryhmään kuuluu 5 geeniä. Myotubulariineista  MTMR ainakin  MTMR2; PTPN9 geenin koodaama PTP-MEG2)

 Täten 81 PTP  entsyymiä  on aktiiveja proteiinifosfataaseja, jotka kykenevät defosforyloimaan  fosfotyrosiinia eli  poistamaan fosfotyrosiinista  (pTyr)  fosfaattia.   

Jos vertaa  proteiinityrosiinikinaasien (PTK)  90 kinaasin suuruiseen  joukkoon , niistä 85 on  katalyyttisesti aktiivia .

Täten  aktiivien tyrosiinifosfataasien (PTP) ja tyrosiinikinaasien (PTK) lukumäärät ovat hyvin samanlaisia. Tästä voisi olettaa että niillä on vertailtavissa olevia substraattispesifisyyksiäkin. 

Kummallakin  entsyymityypillä  on myös  vertailtavissa olevaa  jakaantumistapaa kehossa: yleisestä jakaantumisesta  solutyyppiin rajoittuneeseeen jakaantumiseen siinä määriin, että yksittäiset solut ilmentävät 30%-60%:sti koko PTP-entsyymien kirjoa ja PTK-entsyymien kirjoa. Hermosolut ja vertamuodostavat solut  saattavat ilmentää jopa korkeampia  osuuksia kaikista  proteiinityrosiinifosfataaseista.

Yksittäiset proteiinityrosiinifosfataasit (PTP)  eivät näytä korvaavan toisiansa ja tätä kuvaakin hyvin  raportit  hiirissä esiintyneiden   geenideleetioitten  ainutlaatuisista fenotyypeistä. Esimerkiksi  PTPN22deleetio ( hiirellä PTPN8 deleetio)  aiheuttaa  liiallista T-lymfosyyttimuistisolujen määrän kasvua, pitkittyneitä sekundäärisiä immuunivasteita ja autoimmuniteettia  (Hasegawa et al. 2004). Ihmisellä PTPN22geenin polymorfia korreloi autoimmuunidiabetekseen (Bottini et al. 2004). Muittenkin PTP-geenien mutaatioista seuraa ihmisessä tauteja, mikä  tukee huomiota yksittäisten  proteiinityrosiinifosfataasien  ainutlaatuisista ja tärkeistä toiminnoista..

  Lähde:

Protein Tyrosine Phosphatases in the Human Genome

• Andres Alonso et al.

Open ArchiveDOI:https://doi.org/10.1016/j.cell.2004.05.018

..

 Why So Many PTPs?

The number of genes in the human genome that encode members of the PTP families is higher than anticipated and exceeds the number of genes encoding PTKs (

Manning et al., 2002

). However, a direct comparison of the two numbers, 107 and 90, respectively, is not completely fair: of the 107 PTP genes, 11 are catalytically inactive, 2 dephosphorylate mRNA, and 13 dephosphorylate inositol phospholipids. Thus, 81 PTPs are active protein phosphatases with the ability to dephosphorylate phosphotyrosine. Of the 90 PTKs, 85 are through to be catalytically active. Thus, the numbers of active PTPs and PTKs are very similar and one can therefore assume that they have comparable substrate specificities.

 Both types of enzymes also display comparable tissue distribution patterns, from ubiquitous to cell type restricted, so that individual cells express 30%–60% of the entire complement of PTPs and PTKs. Neuronal and hematopoietic cells may express even higher portions of all PTPs.

 That individual PTPs have nonredundant functions is also well illustrated by the unique phenotypes of many of the reported gene deletions in mice. For example, deletion of PTPN22 (PTPN8 in the mouse) causes excessive expansion of memory T lymphocytes, prolonged secondary immune responses, and autoimmunity (Hasegawa et al., 2004). A polymorphism in the human PTPN22 gene also correlates with autoimmune diabetes in humans (Bottini et al., 2004). Other human diseases caused by mutations in PTP genes (see discussion below) also support the notion that individual PTPs have unique and important functions.