Correct in vivo processing of a chimeric ubiquitin-proapolipoprotein A-I fusion protein in baculovirus-infected insect cells.
The cDNA coding for human proapolipoprotein A-I was expressed as a ubiquitin fusion under the control of the polyhedrin promoter in baculovirus-infected Sf9 Spodoptera frugiperda insect cells. The fusion protein was expressed at high level and was quantitatively cleaved in vivo. The cleaved product was purified and its N-terminal amino acid sequence was established. The data showed that authentic proapolipoprotein A-I has been produced, and thus demonstrated the existence in Spodoptera frugiperda insect cells of a specific ubiquitin hydrolase activity.
Moguilevsky
N
N
University of Brussels, Nivelles, Belgium.
Guillaume
J P
JP
Varsalona
F
F
Bulinckx
M
M
Bollen
A
A
D000595:Amino Acid Sequence; D000818:Animals; D016632:Apolipoprotein A-I; D001054:Apolipoproteins A / Q000096:biosynthesis* / Q000302:isolation & purification; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008075:Lipoproteins, HDL / Q000096:biosynthesis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009036:Moths; D011498:Protein Precursors / Q000096:biosynthesis* / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D014162:Transfection / Q000379:methods; D014452:Ubiquitins / Q000096:biosynthesis*
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A; D018076:DNA, Complementary; D008075:Lipoproteins, HDL; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014452:Ubiquitins