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 language | English |
|---|---|
| Pages (from-to) | 624-632 |
| Number of pages | 9 |
| Journal | Chemistry and Biology |
| Volume | 16 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Jun 26 2009 |
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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 journal › Article
}
TY - JOUR
T1 - Molecular Identification of Albumin and Hsp70 as Cytosolic Anandamide-Binding Proteins
AU - Oddi, Sergio
AU - Fezza, Filomena
AU - Pasquariello, Nicoletta
AU - D'Agostino, Antonella
AU - Catanzaro, Giuseppina
AU - De Simone, Chiara
AU - Rapino, Cinzia
AU - Finazzi-Agrò, Alessandro
AU - Maccarrone, Mauro
PY - 2009/6/26
Y1 - 2009/6/26
N2 - 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.
AB - 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.
KW - CELLBIO
KW - CHEMBIO
UR - http://www.scopus.com/inward/record.url?scp=67649470418&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=67649470418&partnerID=8YFLogxK
U2 - 10.1016/j.chembiol.2009.05.004
DO - 10.1016/j.chembiol.2009.05.004
M3 - Article
C2 - 19481477
AN - SCOPUS:67649470418
VL - 16
SP - 624
EP - 632
JO - Chemistry and Biology
JF - Chemistry and Biology
SN - 1074-5521
IS - 6
ER -