Expression of the N‐ and C‐termini from the Vanilloid Receptor 1 (VR1) as MBP‐fusion proteins for affinity purification and search for interaction partners

• Published in: Journal of neurochemistry Vol. 85; no. s2; p. 31
• Main Authors: Jahnel, R., Goswami, C., Si, H., Dreger, M., Gillen, C., Hucho, F.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01-06-2003
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1046/j.1471-4159.85.s2.20_3.x

The Vanilloid Receptor 1 is a heat‐sensitive, nonselective cation channel expressed by primary sensory neurons involved in nociception. To find and study potential interacting proteins of VR1 we cloned the full N‐ and C‐termini without transmembrane sequences of VR1 into the bacterial expression vector pMALc2x. We here report the expression and purification of the VR1 fragments fused to the maltose binding protein (MBP). In addition, we assessed the subcellular targeting of N‐ and C‐terminal VR1 fragments expressed in the cell line F‐11. This cell line has recently been shown to be particularly suited for the expression of VR1 (1). Preliminary data suggest that the VR1 N‐terminal fragment is targeted to membranes despite the lack of predicted transmembrane regions.