A representation of the cDNA of human FAS DI and DII-DIII plasmids used
in identifying the importance of the ID region in the FAS dimer
formation. The top bars represent, respectively, human FAS monomer and
dimer. The cDNA nucleotide sequences coding for various constituent
activities (speckled bars) and the ID (filled bars) are indicated. The
N- (ID-N) and C-terminal (ID-C) halves of the ID are represented as
gray and black bars, respectively. The human FAS cDNA nucleotide
numbers on the top bar indicate the location of the restriction sites
for BamHI (B), HindIII (H), and
NotI (N), and the boundaries of the coding sequences
present in the recombinant expression plasmids. The cartoon describing
the FAS dimer with two active centers is drawn by using the descriptors
shown for human FAS cDNA shown in the top line. The construction of the
plasmids used for the expression of human FAS recombinant protein
halves containing ID halves is described as before (23, 24), except
that their names have been changed to include the ID sequences present
in them. The constructs lacking ID regions, TRX-hFAS-DI and
TRX-hFAS-DII-III, are described in Materials and
Methods. TRX, thioredoxin (as N-terminal fusion present in the
recombinant protein).