An import-competent precursor of small subunit of ribulose-1,5-bisphosphate carboxylase generated by factor Xa cleavage from a beta-galactosidase fusion expressed in Escherichia coli.
A plasmid-encoding fusion protein interlinked by factor Xa recognition sequence between beta-galactosidase and a precursor of the small subunit of wheat ribulose-1,5-bisphosphate carboxylase has been constructed. The plasmid directed abundant synthesis of the fusion protein in Escherichia coli. The recombinant protein was accumulated in an aggregated form that was associated with the bacterial membranes. A procedure was developed to isolate the fusion protein in a relatively pure and soluble form. Bovine factor Xa cleaved the isolated chimera to generate the complete chloroplast precursor of the small subunit of ribulose-1,5-bisphosphate carboxylase from the fused beta-galactosidase. The cleaved precursor protein was imported into the isolated chloroplasts and processed to yield its mature counterpart.
Evans
A
A
Department of Biochemistry, University College of Wales, Dyfed, United Kingdom.
Liu
Y Y
YY
Kaderbhai
M A
MA
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002736:Chloroplasts / Q000201:enzymology; D004926:Escherichia coli; D015951:Factor Xa / Q000378:metabolism*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D011498:Protein Precursors / Q000096:biosynthesis / Q000302:isolation & purification*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012273:Ribulose-Bisphosphate Carboxylase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D014908:Triticum / Q000201:enzymology / Q000235:genetics; D001616:beta-Galactosidase / Q000096:biosynthesis
D010940:Plant Proteins; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; D015951:Factor Xa; D012273:Ribulose-Bisphosphate Carboxylase