The intron 5-inserted form of rat erythropoietin receptor is expressed as a membrane-bound form.
The cDNA encoding an intron 5-inserted form of the erythropoietin receptor (I5Epo-R) has been cloned from rat. DNA sequence analysis reveals that the insertion of intron 5, which consists of 79 bp, causes a shift in reading frame and results in termination in the region of exon 7. The deduced amino acid sequence is composed of 316 amino acid residues, which is a molecular weight of 34220. To study the function of rat I5Epo-R, we established a Chinese hamster ovary cell line expressing rat I5Epo-R. Western blot analysis and binding studies with 125I-recombinant human erythropoietin showed that the transfected cells expressed rat I5Epo-R with a molecular size of 36 kDa as a membrane-bound form, but not as a soluble form, and had a single class of binding sites with a Kd of 700 pM.
Yamaji
R
R
Department of Applied Biological Chemistry, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan.
Murakami
C
C
Takenoshita
M
M
Tsuyama
S
S
Inui
H
H
Miyatake
K
K
Nakano
Y
Y
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000098:blood supply; D016466:CHO Cells; D006224:Cricetinae; D003432:Cross-Linking Reagents; D004730:Endothelium, Vascular / Q000378:metabolism; D004921:Erythropoietin / Q000378:metabolism; D006801:Humans; D007438:Introns; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D051381:Rats; D017467:Receptors, Erythropoietin / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D012995:Solubility; D014162:Transfection
D003432:Cross-Linking Reagents; D008565:Membrane Proteins; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; D004921:Erythropoietin