TY - CHAP
T1 - Modern methods to investigate the oligomerization of glycoprotein hormone receptors (TSHR, LHR, FSHR)
AU - Bonomi, Marco
AU - Persani, Luca
PY - 2013
Y1 - 2013
N2 - As for other GPCRs, the oligomerization of glycoprotein hormone receptors (GPHRs) appears as critical event for receptor function. By means of modern techniques based on the BRET or FRET principle, GPHR oligomerization has been reported to explain several physiological and pathological conditions. In particular, the presence of oligomers was demonstrated not only in in vitro heterologous systems but also in in vivo tissues, and GPHR homodimerization appears associated with strong negative cooperativity, thus suggesting that one hormone molecule may be sufficient for receptor dimer stimulation. In addition, oligomerization has been reported to occur early during the posttranslational maturation process and to be involved in the dominant negative effect exerted by loss-of-function TSH receptor (TSHR) mutants, that are prevalently retained inside the cell, on the surface expression of wild-type receptors. This molecular mechanism thus explains the dominant inheritance of certain forms of TSH resistance. Here, we provide the description of the methods used in the original BRET, FRET, and HTRF-RET experiments.
AB - As for other GPCRs, the oligomerization of glycoprotein hormone receptors (GPHRs) appears as critical event for receptor function. By means of modern techniques based on the BRET or FRET principle, GPHR oligomerization has been reported to explain several physiological and pathological conditions. In particular, the presence of oligomers was demonstrated not only in in vitro heterologous systems but also in in vivo tissues, and GPHR homodimerization appears associated with strong negative cooperativity, thus suggesting that one hormone molecule may be sufficient for receptor dimer stimulation. In addition, oligomerization has been reported to occur early during the posttranslational maturation process and to be involved in the dominant negative effect exerted by loss-of-function TSH receptor (TSHR) mutants, that are prevalently retained inside the cell, on the surface expression of wild-type receptors. This molecular mechanism thus explains the dominant inheritance of certain forms of TSH resistance. Here, we provide the description of the methods used in the original BRET, FRET, and HTRF-RET experiments.
KW - BRET
KW - Dominant negative effect
KW - FRET
KW - FSHR
KW - Glycoprotein hormone receptors
KW - HTRF-RET
KW - LHR
KW - Negative cooperativity
KW - TSHR
UR - http://www.scopus.com/inward/record.url?scp=84873032959&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84873032959&partnerID=8YFLogxK
U2 - 10.1016/B978-0-12-391862-8.00020-X
DO - 10.1016/B978-0-12-391862-8.00020-X
M3 - Chapter
C2 - 23351750
AN - SCOPUS:84873032959
SN - 9780123918628
VL - 521
T3 - Methods in Enzymology
SP - 367
EP - 383
BT - Methods in Enzymology
ER -