Molecular Identification of Albumin and Hsp70 as Cytosolic Anandamide-Binding Proteins

Sergio Oddi, Filomena Fezza, Nicoletta Pasquariello, Antonella D'Agostino, Giuseppina Catanzaro, Chiara De Simone, Cinzia Rapino, Alessandro Finazzi-Agrò, Mauro Maccarrone

Research output: Contribution to journalArticle

Abstract

The cellular uptake and the intracellular synthesis/degradation of anandamide are crucial steps for controlling its extracellular level and the duration of its activity. Although the biosynthesis and breakdown of anandamide are well understood, little is known about the mechanisms underlying its intracellular transport. Here, we investigated the presence of a potential carrier-mediated trafficking of anandamide within the cytosol, using a biotinylated analog as a tool to catch by affinity chromatography anandamide-interacting proteins. The identity of two of these anandamide-binding proteins, Hsp70 and serum albumin, was determined by mass spectrometry, confirmed by western blotting and confocal microscopy, and further validated through an anandamide-binding assay. These findings suggest that the trafficking of anandamide from the plasma membrane to the internal compartments of a cell occur via a nonvesicular mechanism mediated by cytosolic carriers.

Original languageEnglish
Pages (from-to)624-632
Number of pages9
JournalChemistry and Biology
Volume16
Issue number6
DOIs
Publication statusPublished - Jun 26 2009

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Albumins
Carrier Proteins
Affinity chromatography
Confocal microscopy
Biosynthesis
Cell membranes
anandamide
Affinity Chromatography
Confocal Microscopy
Serum Albumin
Cytosol
Mass spectrometry
Assays
Mass Spectrometry
Western Blotting
Cell Membrane
Degradation
Proteins

Keywords

  • CELLBIO
  • CHEMBIO

ASJC Scopus subject areas

  • Biochemistry
  • Drug Discovery
  • Molecular Biology
  • Clinical Biochemistry
  • Molecular Medicine
  • Pharmacology

Cite this

Molecular Identification of Albumin and Hsp70 as Cytosolic Anandamide-Binding Proteins. / Oddi, Sergio; Fezza, Filomena; Pasquariello, Nicoletta; D'Agostino, Antonella; Catanzaro, Giuseppina; De Simone, Chiara; Rapino, Cinzia; Finazzi-Agrò, Alessandro; Maccarrone, Mauro.

In: Chemistry and Biology, Vol. 16, No. 6, 26.06.2009, p. 624-632.

Research output: Contribution to journalArticle

Oddi, S, Fezza, F, Pasquariello, N, D'Agostino, A, Catanzaro, G, De Simone, C, Rapino, C, Finazzi-Agrò, A & Maccarrone, M 2009, 'Molecular Identification of Albumin and Hsp70 as Cytosolic Anandamide-Binding Proteins', Chemistry and Biology, vol. 16, no. 6, pp. 624-632. https://doi.org/10.1016/j.chembiol.2009.05.004
Oddi S, Fezza F, Pasquariello N, D'Agostino A, Catanzaro G, De Simone C et al. Molecular Identification of Albumin and Hsp70 as Cytosolic Anandamide-Binding Proteins. Chemistry and Biology. 2009 Jun 26;16(6):624-632. https://doi.org/10.1016/j.chembiol.2009.05.004
Oddi, Sergio ; Fezza, Filomena ; Pasquariello, Nicoletta ; D'Agostino, Antonella ; Catanzaro, Giuseppina ; De Simone, Chiara ; Rapino, Cinzia ; Finazzi-Agrò, Alessandro ; Maccarrone, Mauro. / Molecular Identification of Albumin and Hsp70 as Cytosolic Anandamide-Binding Proteins. In: Chemistry and Biology. 2009 ; Vol. 16, No. 6. pp. 624-632.
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