Two isoforms of human membrane-bound alpha Ig resulting from alternative mRNA splicing in the membrane segment.
Antibodies specific for membrane-bound Ig (mIg) but not for their secreted forms would be useful not only for studying the function of mIg but also for modulating B cell activities in vivo. We have proposed that the extracellular portions of the membrane anchor peptides of mIg can be used as antigenic sites for isotype-specific targeting of B cells. Clones containing the genes of human Ig alpha 1 or alpha 2 subclasses were isolated from a genomic DNA library. The gene segments encoding the membrane peptides and their flanking regions were amplified by polymerase chain reaction, subcloned into plasmid pUC19, and the DNA sequences were determined. Human alpha 1 and alpha 2 genes, like murine alpha gene, each has only one membrane exon. The sequences of the human alpha 1 and alpha 2 genes are almost identical in the membrane peptide-coding region. The mRNA from a human mIgA-expressing B cell line, DAKIKI, was isolated, its cDNA prepared, and the segments spanning the membrane peptide-coding region and a part of the constant domain 3 amplified by polymerase chain reaction. DNA sequences revealed that there are two isoforms of alpha 1-chain, resulting from the alternative splicing of the third constant domain of H chain to two acceptor sites in the membrane exon. One isoform has a segment of 32 and the other 26 amino acid residues in the extracellular portion of the membrane peptide. These segments may serve as isotype-specific antigenic epitopes for antibody targeting of mIgA-bearing B cells.
Yu
L M
LM
Tanox Biosystems, Inc., Houston, TX 77025.
Peng
C
C
Starnes
S M
SM
Liou
R S
RS
Chang
T W
TW
D001483:Base Sequence; D004247:DNA / Q000032:analysis; D005803:Genes, Immunoglobulin; D006801:Humans; D007132:Immunoglobulin Isotypes / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D011947:Receptors, Antigen, B-Cell / Q000235:genetics*
D007132:Immunoglobulin Isotypes; D012333:RNA, Messenger; D011947:Receptors, Antigen, B-Cell; D004247:DNA