The isolation and in situ location of adligin: the microtubule cross-linking protein from Caenorhabditis elegans.
Microtubules isolated from the nematode Caenorhabditis elegans contain long stretches of periodic cross-links formed by microtubule-associated proteins (MAPs). These cross-links are 5.7 nm long, 3 nm wide, and occur at one tubulin dimer (8-nm) intervals along the walls of microtubules (Aamodt, E., and J. Culotti, 1986. J. Cell Biol. 103:23-31). The structural protein of the cross-links was isolated from the MAPs by centrifugation and exclusion chromatography. The cross-links were formed exclusively from the most prevalent MAP, a 32,000 mol wt protein. We suggest the name adligin for this MAP. Adligin eluted from the exclusion column at 33,000 mol wt indicating that it was a monomer in solution. Antibodies were made against the purified adligin and affinity purified. The affinity-purified antibodies were used to locate adligin in situ and to determine its distribution relative to that of tubulin by the use of double label immunofluorescence. The anti-adligin antibodies labeled a fibrous network in the cytoplasm of most cells of C. elegans. Neurons were labeled especially well. This labeling pattern was similar to the labeling pattern obtained with antitubulin, but anti-adligin labeled some granules in the gut that were not labeled with antitubulin. These results suggest that adligin may be part of the interphase microtubule network in C. elegans.
Aamodt
E
E
Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208.
Holmgren
R
R
Culotti
J
J
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002107:Caenorhabditis / Q000032:analysis* / Q000254:growth & development; D029742:Caenorhabditis elegans Proteins; D003460:Crystallization; D005455:Fluorescent Antibody Technique; D007399:Interphase; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D008870:Microtubules / Q000032:analysis*; D008970:Molecular Weight; D009474:Neurons / Q000032:analysis; D014404:Tubulin / Q000032:analysis
D000906:Antibodies; D029742:Caenorhabditis elegans Proteins; D008869:Microtubule-Associated Proteins; D014404:Tubulin; C058791:adligin protein, C elegans