Expression of chimeric monomer and dimer proteins on the plasma membrane of mammalian cells.
Targeting of proteins to the plasma membrane of cells may be useful for vaccine development, tissue engineering, genetic research, bioseparations, and disease treatment. The ability of different transmembrane domains (TM) to direct a reporter protein (human alpha-feto protein, AFP) to the surface of mammalian cells was examined. High surface expression was achieved with chimeric proteins composed of AFP and the TM and cytosolic tail of murine B7-1 (AFP-B7) as well as with AFP containing a GPI-anchor from decay-accelerating factor (AFP-DAF). Lower surface expression of AFP was observed when the TM of human platelet-derived growth factor receptor or the human asialoglycoprotein receptor H1 subunit were employed. Introduction of the hinge-CH2-CH3 region of human IgG (gamma1 domain) between AFP and TM allowed efficient formation of disulfide-linked dimers. Surface expression of AFP-gamma1-B7 dimers was impaired compared to AFP-B7 whereas AFP-gamma1-DAF dimers were efficiently targeted to the surface. Accumulation of chimeric proteins on the cell surface did not correlate with the level of protein expression. This study demonstrates that high levels of monomeric and dimeric proteins can be targeted to the cell membrane of mammalian cells by proper selection of TM.
Chou
W C
WC
Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan.
Liao
K W
KW
Lo
Y C
YC
Jiang
S Y
SY
Yeh
M Y
MY
Roffler
S R
SR
D000818:Animals; D037263:Asialoglycoprotein Receptor; D018122:B7-1 Antigen / Q000235:genetics; D018958:CD55 Antigens / Q000235:genetics; D002460:Cell Line; D017931:DNA Primers; D019281:Dimerization; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D010982:Platelet-Derived Growth Factor / Q000235:genetics; D011487:Protein Conformation; D011956:Receptors, Cell Surface / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D014162:Transfection; D000509:alpha-Fetoproteins / Q000378:metabolism
D037263:Asialoglycoprotein Receptor; D018122:B7-1 Antigen; D018958:CD55 Antigens; D017931:DNA Primers; D008565:Membrane Proteins; D010982:Platelet-Derived Growth Factor; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D000509:alpha-Fetoproteins