Functional expression and characterization of the C. elegans G-protein-coupled FLP-2 Receptor (T19F4.1) in mammalian cells and yeast

被引:12
|
作者
Larsen, Martha J. [1 ]
Lancheros, Elizabeth Ruiz [2 ]
Williams, Tracey [1 ]
Lowery, David E. [1 ]
Geary, Timothy G. [1 ]
Kubiak, Teresa M. [1 ]
机构
[1] Pfizer Anim Hlth Discovery Res, Vet Med Discovery Res, Kalamazoo, MI 49001 USA
[2] McGill Univ, Inst Parasitol, Ste Anne De Bellevue, PQ H9X 3V9, Canada
关键词
FLP; Caenorhabditis elegans; flp2; Neuropeptide; CAENORHABDITIS-ELEGANS; SACCHAROMYCES-CEREVISIAE; DROSOPHILA-MELANOGASTER; NERVOUS-SYSTEM; GENE FAMILIES; NEUROPEPTIDE; PEPTIDES; IDENTIFICATION; CLONING; NEUROTRANSMITTERS;
D O I
10.1016/j.ijpddr.2012.10.002
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
Profound neuropeptide diversity characterizes the nematode nervous system, but it has proven challenging to match neuropeptide G protein-coupled receptors (GPCR) with their cognate ligands in heterologous systems. We have expressed the Caenorhabditis elegans GPCR encoded in the locus T19F4.1, previously matched with FMRFamide-like peptides encoded on the flp-2 precursor gene, in mammalian cells and in the yeast Saccharomyces cerevisiae. Pharmacological characterization revealed that the receptor is potently activated by flp-2 peptides in CHO cells (similar to 10 nM EC50) and in yeast (similar to 100 nM EC50), signaling through a G(q)alpha pathway in each system. The yeast GPCR expression system provides a robust assay for screening for agonists of the flp-2 receptor and is the target of an ongoing high-throughput screening exercise. (C) 2012 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:1 / 7
页数:7
相关论文
共 30 条
  • [1] Successful expression of a functional yeast G-protein-coupled receptor (Ste2) in mammalian cells
    Yin, DZ
    Gavi, S
    Shumay, E
    Duell, K
    Konopka, JB
    Malbon, CC
    Wang, HY
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2005, 329 (01) : 281 - 287
  • [2] Expression of a mammalian olfactory G-protein-coupled receptor in C-elegans
    Milani, N
    Guarin, E
    Renfer, E
    Nef, P
    Andres-Barquin, PJ
    MOLECULAR BIOLOGY OF THE CELL, 2002, 13 : 123A - 124A
  • [3] Functional annotation of the putative orphan Caenorhabditis elegans G-protein-coupled receptor C10C6.2 as a FLP15 peptide receptor
    Kubiak, TM
    Larsen, MJ
    Zantello, MR
    Bowman, JW
    Nulf, SC
    Lowery, DE
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (43) : 42115 - 42120
  • [4] CLONING - EXPRESSION, AND CHARACTERIZATION OF A G-PROTEIN-COUPLED, BOVINE PARATHYROID CA2+ RECEPTOR
    BROWN, EM
    GAMBA, G
    RICARDI, D
    LOMBARDI, M
    BUTTERS, R
    KIFOR, O
    HEDIGER, M
    HEBERT, SC
    JOURNAL OF BONE AND MINERAL RESEARCH, 1993, 8 : S147 - S147
  • [5] Molecular characterization of two G protein-coupled receptor splice variants as FLP2 receptors in Caenorhabditis elegans
    Mertens, I
    Meeusen, T
    Janssen, T
    Nachman, R
    Schoofs, L
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2005, 330 (03) : 967 - 974
  • [6] Functional characterization of the putative orphan neuropeptide G-protein coupled receptor C26F1.6 in Caenorhabditis elegans
    Mertens, I
    Vandingenen, A
    Meeusen, T
    Janssen, T
    Luyten, W
    Nachman, RJ
    De Loof, A
    Schoofs, L
    FEBS LETTERS, 2004, 573 (1-3) : 55 - 60
  • [7] Morphological characterization of rat Mas-related G-protein-coupled receptor C and functional analysis of agonists
    Hager, U. A.
    Hein, A.
    Lennerz, J. K. M.
    Zimmermann, K.
    Neuhuber, W. L.
    Reeh, P. W.
    NEUROSCIENCE, 2008, 151 (01) : 242 - 254
  • [8] Investigation of the neuronal expression pattern of the G protein-coupled receptor FSHR-1 in C. elegans.
    Nettesheim, E. J.
    Cherry, K. L.
    Emch, D.
    Kowalski, J. R.
    MOLECULAR BIOLOGY OF THE CELL, 2018, 29 (26)
  • [9] FMRFamide-like peptides encoded on the flp-18 precursor gene activate two Isoforms of the orphan Caenorhabditis elegans G-protein-coupled receptor Y58G8A.4 heterologously expressed in mammalian cells
    Kubiak, Teresa M.
    Larsen, Martha J.
    Bowman, Jerry W.
    Gearyt, Timothy G.
    Lowery, David E.
    BIOPOLYMERS, 2008, 90 (03) : 339 - 348
  • [10] Cloning, Expression, Purification and Characterization of Oligomeric States of the Native 5HT2A G-Protein-Coupled Receptor
    Mahesh, Gopa
    Jaiswal, Priyank
    Dey, Sandip
    Sengupta, Jayati
    Mukherjee, Sujoy
    PROTEIN AND PEPTIDE LETTERS, 2018, 25 (04): : 390 - 397