Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 105 - 107 |
| Number of pages | 3 |
| Journal | Annals of the New York Academy of Sciences |
| Volume | 1160 |
| DOIs | |
| Publication status | Published - 2009 |
Cite this
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Addition of a carboxy-terminal green fluorescent protein does not alter the binding and signaling properties of relaxin family peptide receptor 3. / van der Westhuizen, Emma Therese; Wade, John D; Sexton, Patrick M; Summers, Roger James.
In: Annals of the New York Academy of Sciences, Vol. 1160, 2009, p. 105 - 107.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Addition of a carboxy-terminal green fluorescent protein does not alter the binding and signaling properties of relaxin family peptide receptor 3
AU - van der Westhuizen, Emma Therese
AU - Wade, John D
AU - Sexton, Patrick M
AU - Summers, Roger James
PY - 2009
Y1 - 2009
N2 - 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 receptor function. This study demonstrated that addition of a carboxy-terminal fusion protein to RXFP3 did not alter the binding or signaling properties of RXFP3, making RXFP3-GFP(2) a useful tool for future receptor localization and trafficking studies.
AB - 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 receptor function. This study demonstrated that addition of a carboxy-terminal fusion protein to RXFP3 did not alter the binding or signaling properties of RXFP3, making RXFP3-GFP(2) a useful tool for future receptor localization and trafficking studies.
UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19416168
U2 - 10.1111/j.1749-6632.2008.03818.x
DO - 10.1111/j.1749-6632.2008.03818.x
M3 - Article
VL - 1160
SP - 105
EP - 107
JO - Annals of the New York Academy of Sciences
JF - Annals of the New York Academy of Sciences
SN - 0077-8923
ER -