C6 epitope expression by an unrelated antisense cDNA clone: an inadvertent surface-simulation mimotope.
A cDNA clone which directs the expression of a fusion protein reacting with anti-C6 antibodies has been isolated and sequenced. A synthetic peptide corresponding to the 14 C-terminal residues of the expressed protein elicited the production of antibodies which are specific for native C6, confirming the presence of a C6 epitope on the expressed protein. However, analysis of the intron-exon boundaries of a corresponding genomic clone revealed that the expression clone is in antisense orientation, and is therefore not C6 cDNA. Comparison of the sequences of the expression clone and expressed protein with those for C6 have not demonstrated any significant sequence homology. It is therefore apparent that what has been cloned is a mimotope for C6 which includes in its continuous sequence an epitope that is conformational in C6 and not represented as a continuous sequence in the C6 molecule. Although this was not the purpose of the investigation, these results confirm that screening random expression libraries with antibodies may be an alternative to the synthetic peptide approach to obtain mimotopes reacting with particular antibodies.
Hobart
M J
MJ
Molecular Immunopathology Unit, M.R.C. Centre, Cambridge, U.K.
Fernie
B A
BA
Stanley
K K
KK
Lachmann
P J
PJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D003183:Complement C6 / Q000096:biosynthesis* / Q000235:genetics; D016373:DNA, Antisense; D000939:Epitopes / Q000096:biosynthesis* / Q000235:genetics; D005091:Exons; D015870:Gene Expression; D015723:Gene Library; D015698:Genomic Library; D006801:Humans; D007438:Introns; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010455:Peptides / Q000138:chemical synthesis; D012333:RNA, Messenger / Q000378:metabolism
D003183:Complement C6; D016373:DNA, Antisense; D000939:Epitopes; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D012333:RNA, Messenger