Expression and purification of the D4 region of PLD1 and characterization of its interaction with PED-PEA15 (Articolo in rivista)

Type

• Articolo in rivista (Classe)
• Prodotto della ricerca (Classe)

Label

• Expression and purification of the D4 region of PLD1 and characterization of its interaction with PED-PEA15 (Articolo in rivista) (literal)

Anno

• 2008-01-01T00:00:00+01:00 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi

• 10.1016/j.pep.2008.02.012 (literal)

Alternative label

• Viparelli F; Doti N; Sandomenico A; Marasco D; Dathan NA; Miele C; Beguinot F; Monti SM; Ruvo M. (2008)
  Expression and purification of the D4 region of PLD1 and characterization of its interaction with PED-PEA15
  in Protein expression and purification (Print)
  (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

• Viparelli F; Doti N; Sandomenico A; Marasco D; Dathan NA; Miele C; Beguinot F; Monti SM; Ruvo M. (literal)

Pagina inizio

• 302 (literal)

Pagina fine

• 308 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

• 59 (literal)

Rivista

• Protein expression and purification (Rivista)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#descrizioneSinteticaDelProdotto

• Pubblicazione su rivista internazionale (literal)

Note

• ISI Web of Science (WOS) (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

• CNR, IBB, I-80134 Naples, Italy Univ Naples Federico 2, Dipartimento Sci Biol, I-80134 Naples, Italy Seconda Univ Napoli, Dipartimento Biochim & Biofis, I-80138 Naples, Italy CNR, IEOS, I-80131 Naples, Italy (literal)

Titolo

• Expression and purification of the D4 region of PLD1 and characterization of its interaction with PED-PEA15 (literal)

Abstract

• PLD's (Phospholipases D) are ubiquitously expressed proteins involved in many transphosphatidylation reactions. They have a bi-lobed structure composed by two similar domains which at their interface reconstitute the catalytic site through the association of the two conserved HxKx(4)DX(6)GSxN motifs. PLD1 interacts with the small phosphoprotein PED-PEA15 by an unknown mechanism that, by enhancing PLD1 stability, apparently increases its enzymatic activity; the minimum interacting region of PLD1 was previously identified as spanning residues 712-1074 (D4 region). Since the D4/PED-PEA15 interaction has been claimed to be one of the multiple molecular events that can trigger type 2 diabetes, we purified the two recombinant proteins to study in vitro this binding by both ELISA and SPR techniques. Whilst PED-PEA15 was easily expressed and purified, expression of recombinant D4 was more problematic and only the fusion protein with Thioredoxin A and a six Histidine Tag (Trx-HiS(6)-D4) demonstrated sufficient stability for further characterization. We have found that Trx-His(6)-D4 is present as two different oligorneric forms, though only the monomeric variant is able to interact with PED-PEA15. All these findings may have important implications for both the mechanisms of phospholipase activity and PED-PEA15 regulative functions. (C) 2008 Elsevier Inc. All rights reserved (literal)

Prodotto di

• Institute of biostructure and bioimaging (IBB) (Istituto)
• Studio dell'espressione e della funzione di geni e proteine coinvolte nel diabete di tipo 2 (SV.P16.001.002) (Modulo)
• Istituto per l'endocrinologia e l'oncologia \"Gaetano Salvatore\" (IEOS) (Istituto)
• Nuove molecole ad azione farmacologica per il trattamento del diabete di tipo II (PM.P01.005.004) (Modulo)

Autore CNR

• FRANCESCO BEGUINOT (Unità di personale esterno)
• NINA ALAYNE DATHAN (Persona)
• MENOTTI RUVO (Unità di personale interno)
• CLAUDIA MIELE (Persona)
• SIMONA MARIA MONTI (Unità di personale interno)

Insieme di parole chiave

• Keywords of "Expression and purification of the D4 region of PLD1 and characterization of its interaction with PED-PEA15" (Insieme di parole chiave)

Incoming links:

Autore CNR di

• FRANCESCO BEGUINOT (Unità di personale esterno)
• SIMONA MARIA MONTI (Unità di personale interno)
• NINA ALAYNE DATHAN (Persona)
• MENOTTI RUVO (Unità di personale interno)
• CLAUDIA MIELE (Persona)

Prodotto

• Istituto per l'endocrinologia e l'oncologia \"Gaetano Salvatore\" (IEOS) (Istituto)
• Institute of biostructure and bioimaging (IBB) (Istituto)
• Studio dell'espressione e della funzione di geni e proteine coinvolte nel diabete di tipo 2 (SV.P16.001.002) (Modulo)
• Nuove molecole ad azione farmacologica per il trattamento del diabete di tipo II (PM.P01.005.004) (Modulo)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#rivistaDi

• Protein expression and purification (Rivista)

Insieme di parole chiave di

• Keywords of "Expression and purification of the D4 region of PLD1 and characterization of its interaction with PED-PEA15" (Insieme di parole chiave)