Fluorescent ligands, antibodies, and proteins for the study of receptors

被引:107
作者
Daly, CJ [1 ]
McGrath, JC [1 ]
机构
[1] Univ Glasgow, Fac Biomed & Life Sci, Div Neurosci & Biomed Syst, Glasgow G12 8QQ, Lanark, Scotland
关键词
fluorescence; fluorescence resonance energy transfer; green fluorescent protein; antibodies; receptors; imaging;
D O I
10.1016/j.pharmthera.2003.08.001
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Fluorescent molecules bound to receptors can show their location and, if binding is reversible, can provide pharmacological information such as affinity and proximity between interacting molecules. The spatial precision offered by visualisation transcends the diverse localisation and low molecular concentration of receptor molecules. Consequently, the relationships between receptor location and function and life cycles of receptors have become better understood as a result of fluorescent labeling. Each of these aspects contributes new insights to drug action and potential new targets. The relationships between spatial distribution of receptor and function are largely unknown. This is particularly apparent for native receptors expressed in their normal host tissues where communication between heterogeneous cell types influences receptor distribution and function. In cultured cell systems, particularly for G-protein-coupled receptors (GPCR), fluorescence-based methods have enabled the visualisation of the cycle of agonist-stimulated receptor clustering, endocytic internalisation to the perinuclear region, degradation of the receptor-ligand complex, and recycling back to the surface membrane. Using variant forms of green fluorescent protein (GFP), antibodies, or fluorescent ligands, it is possible to detect or visualise the formation of oligomeric receptor complexes. Careful selection of fluorescent molecules based on their spectral properties enables resonance energy transfer and multilabel visualisation with colocalisation studies. Fluorescent agonist and antagonist ligands are now being used in parallel with GFP to study receptor cycling in live cells. This review covers how labeling and visualisation technologies have been applied to the study of major pharmacologically important receptors and illustrates this by giving examples of recent techniques that have relied on GFP, antibodies, or fluorescent ligands alone or in combination for the purpose of studying GPCR. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:101 / 118
页数:18
相关论文
共 119 条
  • [11] CATALYZED REPORTER DEPOSITION, A NOVEL METHOD OF SIGNAL AMPLIFICATION - APPLICATION TO IMMUNOASSAYS
    BOBROW, MN
    HARRIS, TD
    SHAUGHNESSY, KJ
    LITT, GJ
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 1989, 125 (1-2) : 279 - 285
  • [12] Immunofluorescent imaging of β1- and β2-adrenergic receptors in rat kidney
    Boivin, V
    Jahns, R
    Gambaryan, S
    Ness, W
    Boege, F
    Lohse, MJ
    [J]. KIDNEY INTERNATIONAL, 2001, 59 (02) : 515 - 531
  • [13] The use of resonance energy transfer in high-throughput screening: BRET versus FRET
    Boute, N
    Jockers, R
    Issad, T
    [J]. TRENDS IN PHARMACOLOGICAL SCIENCES, 2002, 23 (08) : 351 - 354
  • [14] Brahmadevara N, 2002, BRIT J PHARMACOL, V137
  • [15] Intermittent ATP release from nerve terminals elicits focal smooth muscle Ca2+ transients in mouse vas deferens
    Brain, KL
    Jackson, VM
    Trout, SJ
    Cunnane, TC
    [J]. JOURNAL OF PHYSIOLOGY-LONDON, 2002, 541 (03): : 849 - 862
  • [16] BROWN GL, 1957, J PHYSIOL-LONDON, V138, P81, DOI 10.1113/jphysiol.1957.sp005839
  • [17] Mast cell degranulating peptide binds to RBL-2H3 mast cell receptors and inhibits IgE binding
    Buku, A
    Price, JA
    Mendlowitz, M
    Masur, S
    [J]. PEPTIDES, 2001, 22 (12) : 1993 - 1998
  • [18] DALY C, 2002, PHARMACOLOGIST S1, V44
  • [19] A knockout approach indicates a minor vasoconstrictor role for vascular α1B-adrenoceptors in mouse
    Daly, CJ
    Deighan, C
    McGee, A
    Mennie, D
    Ali, Z
    McBride, M
    McGrath, JC
    [J]. PHYSIOLOGICAL GENOMICS, 2002, 9 (02) : 85 - 91
  • [20] Daly CJ, 1998, J PHARMACOL EXP THER, V286, P984