TY - JOUR
T1 - Functional analysis of five endothelin-B receptor mutations found in human hirschsprung disease patients
AU - Abe, Yoichiro
AU - Sakurai, Takeshi
AU - Yamada, Takahisa
AU - Nakamura, Toshiaki
AU - Yanagisawa, Masashi
AU - Goto, Katsutoshi
N1 - Funding Information:
This work was supported in part by research grants from the University of Tsukuba Project Research, the Ministry of Education, Science and Culture of Japan and the Japan Society for the Promotion of Science (JSPS-RFTF96I00202). M.Y. is an Investigator of the Howard Hughes Medical Institute. We thank Dr. Robert J. Lefkowitz for donating pcDV1R α1B, Dr. Ikuo Nishimoto for critical reading of the manuscript, and Dr. Wendy Gray for help with preparation of the manuscript.
PY - 2000/8/28
Y1 - 2000/8/28
N2 - Several missense mutations of the endothelin-B receptor (EDNRB) associated with Hirschsprung disease have recently been identified. Five mutated EDNRB (A183G, W276C, R319W, M374I and P383L) cDNAs were transiently expressed in several cell lines to examine the effects of these mutations. Ligand-receptor binding experiments demonstrated that all mutants examined here accept endothelins with a high affinity. Especially, the affinity of endothelins to P383L was increased. However, the number of binding sites of A183G, W276C and P383L was markedly decreased. The subcellular localization of these mutant receptors was the same as that of wild-type EDNRB, whereas the amount of protein of each mutant receptor was decreased. All mutant receptors were impaired in intracellular Ca2+ mobilization. These findings indicate that these missense mutations result in loss of function of EDNRB, and may provide the molecular pathological basis of Hirschsprung disease in some individuals. (C) 2000 Academic Press.
AB - Several missense mutations of the endothelin-B receptor (EDNRB) associated with Hirschsprung disease have recently been identified. Five mutated EDNRB (A183G, W276C, R319W, M374I and P383L) cDNAs were transiently expressed in several cell lines to examine the effects of these mutations. Ligand-receptor binding experiments demonstrated that all mutants examined here accept endothelins with a high affinity. Especially, the affinity of endothelins to P383L was increased. However, the number of binding sites of A183G, W276C and P383L was markedly decreased. The subcellular localization of these mutant receptors was the same as that of wild-type EDNRB, whereas the amount of protein of each mutant receptor was decreased. All mutant receptors were impaired in intracellular Ca2+ mobilization. These findings indicate that these missense mutations result in loss of function of EDNRB, and may provide the molecular pathological basis of Hirschsprung disease in some individuals. (C) 2000 Academic Press.
KW - Endothelin
KW - Endothelin-B receptor
KW - Hirschsprung disease
KW - Intracellular Ca mobilization
KW - Missense mutations
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U2 - 10.1006/bbrc.2000.3291
DO - 10.1006/bbrc.2000.3291
M3 - Article
C2 - 10964697
AN - SCOPUS:0034726747
SN - 0006-291X
VL - 275
SP - 524
EP - 531
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -