Oligomer formation of histamine H2 receptors expressed in Sf9 and COS7 cells.
A histamine H2 receptor, which had been mutated at its glycosylation site and tagged at its N-terminus with an HA tag (HA-H2 receptor), was expressed in Sf9 cells and COS7 cells. Immunoprecipitation and immunoblotting of HA-H2 receptors with alphaHA antibody revealed four bands of 31.5 +/- 2.5 kDa, 59.0 +/- 6.0 kDa, 80.5 +/- 4.5 kDa and 120 kDa. These bands were also detected by immunoblot using anti-H2 receptor serum (C-terminus). In addition, H2 receptors without the HA-tag coimmunoprecipitated with HA-tagged H2 receptors devoid of the 51 C-terminal amino acids, via immunoprecipitation with alphaHA antibody, when the two receptors were coexpressed. These results suggest that H2 receptors are present as receptor oligomers and that the C-terminal portion is not involved in the formation of these oligomers.
Fukushima
Y
Y
Third Department of Internal Medicine, Faculty of Medicine, University OF TOKYO, JAPAN. FUKUSIMA-TKY@UMIN.U-TOKYO.AC.JP
Asano
T
T
Saitoh
T
T
Anai
M
M
Funaki
M
M
Ogihara
T
T
Katagiri
H
H
Matsuhashi
N
N
Yazaki
Y
Y
Sugano
K
K
D000818:Animals; D019556:COS Cells; D003001:Cloning, Molecular; D006388:Hemagglutinins / Q000235:genetics; D015151:Immunoblotting; D008970:Molecular Weight; D011233:Precipitin Tests; D011970:Receptors, Histamine H2 / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D018411:Spodoptera / Q000166:cytology / Q000235:genetics / Q000378:metabolism
D006388:Hemagglutinins; D011970:Receptors, Histamine H2