Conference Proceedings

Addition of a Carboxy-Terminal Green Fluorescent Protein Does Not Alter the Binding and Signaling Properties of Relaxin Family Peptide Receptor 3

Emma T van der Westhuizen, John D Wade, Patrick M Sexton, Roger J Summers, GD BryantGreenwood (ed.), CA Bagnell (ed.), RAD Bathgate (ed.)

RELAXIN AND RELATED PEPTIDES: FIFTH INTERNATIONAL CONFERENCE | WILEY-BLACKWELL | Published : 2009

Abstract

The relaxin family peptide receptor 3 (RXFP3) is the cognate receptor for the neuropeptide relaxin-3. RXFP3 was tagged at the carboxy-terminus with a variant of the green fluorescent protein (GFP(2)) for use in receptor localization studies. RXFP3-GFP(2) was examined to ensure it retained binding and signaling properties similar to untagged RXFP3. Competition for [(125)I]INSL5/H3 relaxin chimera binding to RXFP3 and RXFP3-GFP(2) indicated that the carboxy-terminal tag did not affect receptor binding or receptor internalization. RXFP3-GFP(2) activated ERK1/2 with a similar potency to RXFP3 when transiently expressed in CHO-K1 or HEK293T cells, suggesting that the GFP(2) tag did not affect rec..

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