N-terminal and cDNA characterization of murine lymphocyte antigen Ly-6C.2.
The Ly-6C.2 molecule was purified from K36 tumor cells by affinity chromatography and gel filtration. The electrophoretically homogeneous preparation, with m.w. 15,000, was tested with a panel of antibodies that confirmed the presence of the LY-6C.2 epitope. An N-terminal sequence of 39 amino acids was obtained showing 59% homology with the corresponding portion of the Ly-6A.2 polypeptide. Based on the least homologous (29%) 14 amino acid segment, an oligonucleotide probe was constructed, and Ly-6C.2 cDNA was cloned from a BW5147 cDNA library. A 794-base pair cDNA containing the entire coding region had 82% homology with Ly-6A.2 cDNA. The encoded polypeptide sequence of 131 amino acids containing a perfect correlation with the N-terminal sequence data was 63% homologous with that of Ly-6A.2. The greatest homology was in the leader, first 16 N-terminal and last 39 C-terminal amino acids. The latter are likely to be important in determining the attachment of glycophosphatidylinositol. Despite results indicating fewer disulfide constraints in the Ly-6C molecule, the predicted sequence contains 10 cysteine residues nearly perfectly matched with those predicted in Ly-6A.
Palfree
R G
RG
Sloan-Kettering Institute for Cancer Research, New York, NY 10021.
Sirlin
S
S
Dumont
F J
FJ
Hämmerling
U
U
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000950:Antigens, Ly / Q000235:genetics*; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D005796:Genes; D051379:Mice; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012689:Sequence Homology, Nucleic Acid
D000911:Antibodies, Monoclonal; D000950:Antigens, Ly; D004247:DNA