Expression of the Rh-related glycoprotein (Rh50).
The Rh50 glycoprotein is suspected of being involved in Rh antigen expression. We prepared Rh50 cDNA from a human bone marrow library by polymerase chain reaction and then subcloned this cDNA into various vectors. The vector containing Rh50 cDNA produced a 30-kDa nonglycosylated form of Rh50 in a rabbit reticulocyte lysate system and produced partially glycosylated Rh50 (32 kDa) when microsomes were added to the system. COS-1 cells transiently transfected with the vector containing Rh50 cDNA produced partially glycosylated Rh50 (32 kDa) recognized by a Rh50-specific antibody. Surface expression of Rh50 in K562 cells was also detected by flow cytometry using mouse monoclonal antibody (2D10) specific to Rh50. Partially glycosylated Rh50 (32 kDa) was again isolated from the lysates of K562 cells metabolically labeled with [35S]-methionine or [3H]-mannose using anti-Rh50 antisera. These systems (K562 and COS-1 cells) should prove useful for studying the transport of Rh proteins within the cell and the necessary components needed for Rh antigenicity at the cell surface.
Suyama
K
K
Lindsley F. Kimball Research Institute, New York Blood Center, New York, N.Y. 10021, USA. ksuyama@nybc.org
Lunn
R
R
Smith
B L
BL
Haller
S
S
D000818:Animals; D001798:Blood Proteins / Q000096:biosynthesis* / Q000235:genetics; D019556:COS Cells; D018076:DNA, Complementary / Q000032:analysis; D005784:Gene Amplification; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D020014:K562 Cells; D008562:Membrane Glycoproteins; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012204:Rh-Hr Blood-Group System / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection