Is tubulin the sole antigen recognized by a putative anti-bursicon antibody?
A 56-kDa polypeptide suspected to be the tanning hormone 'bursicon' was analyzed using the monoclonal antibody (mAb) 01C10 of Song and Ma. We studied the beetle Tenebrio molitor, for which data on bursicon have been recently published. After purification by two-dimensional gel electrophoresis of brain proteins, the immunoreactive 56-kDa polypeptide was trypsinated and microsequenced. The obtained sequences revealed a high homology with alpha- and beta-tubulins. In a complementary study, immunoreactive clones were isolated, using the 01C10 mAb, from a library in expression vector obtained from Drosophila melanogaster head cDNAs. Again, the isolated clones were found, after cDNA sequencing, to correspond to tubulin. Our results suggest that, although the 01C10 mAb could possibly still have a great affinity for a polypeptide present in very low quantities in a few brain neurosecretory cells, it also proved to have an artefactual affinity for a 56-kDa polypeptide, identified as tubulin, which is not involved in tanning control.
D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004330:Drosophila / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D015723:Gene Library; D007447:Invertebrate Hormones / Q000276:immunology* / Q000502:physiology; D017422:Sequence Analysis, DNA; D013712:Tenebrio / Q000502:physiology*; D013997:Time Factors; D014404:Tubulin / Q000276:immunology* / Q000378:metabolism / Q000502:physiology