Primary structure of Xenopus laevis S10, a ribosomal protein that cross-reacts with antibodies to immunoglobulin light chains.
Screening of a cDNA expression library from Xenopus laevis splenocytes with purified antibodies to Xenopus immunoglobulin light chains unexpectedly led to the isolation of a clone with an insert whose deduced amino acid sequence is similar to that of a segment of a protein, S10, from the small (40S) subunit of rat ribosomes. A clone containing an insert encoding the corresponding complete protein was isolated from another cDNA library by nucleic acid hybridization. The deduced amino acid sequence of this insert is 94% identical to that of rat S10; no similarity to immunoglobulin sequences could be discerned. The reactivity of the anti-light chain antibodies with the putative Xenopus S10 facilitated the purification of the protein, by high-pressure liquid chromatography, from the 40S subunit of Xenopus ribosomes. Amino-terminal sequence analysis established the identity of the ribosomal protein with the protein encoded by the cDNA insert. To explore the basis for this unexpected cross-reaction, an "antibody transfer" experiment was carried out. Antibodies to Xenopus light chains were adsorbed to Xenopus S10 on a nitrocellulose strip, which was incubated with another strip containing separated heavy and light chains from Xenopus IgM. Antibodies migrated from the strip carrying S10 to the light chains, but not the heavy chains, on the second strip. These results suggest that this unexpected cross-reaction is due to the sharing of one or more epitopes by Xenopus immunoglobulin light chains and the ribosomal protein, S10.
Sarrowa
J
J
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Steiner
L A
LA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D004275:DNA, Ribosomal / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D015723:Gene Library; D015151:Immunoblotting; D007147:Immunoglobulin Light Chains / Q000276:immunology*; D008969:Molecular Sequence Data; D051381:Rats; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D004275:DNA, Ribosomal; D000939:Epitopes; D007147:Immunoglobulin Light Chains; D012269:Ribosomal Proteins; C029537:ribosomal protein S10