Tektins as structural determinants in basal bodies.
Tektins, present as three equimolar 47-55 kDa protein components, form highly insoluble protofilaments that are integral to the junctional region of outer doublet microtubules in cilia and flagella. To identify and quantify tektins in other compound microtubules such as centrioles or basal bodies, a rabbit antiserum was raised against tektin filaments isolated from Spisula solidissima (surf clam) sperm flagellar outer doublets and affinity-purified with nitrocellulose blot strips of tektins resolved by SDS- or SDS-urea-PAGE. These antibodies recognized analogous tektins in axonemes of organisms ranging from ctenophores to higher vertebrates. Quantitative immunoblotting established that outer doublet tektins occur in a 1:17 weight ratio to tubulin. Cilia and basal apparatuses were prepared from scallop gill epithelial cells; cilia and deciliated cells were prepared from rabbit trachea. Tektins were detected by immunoblotting in basal body-enriched preparations while tektins were localized to individual basal bodies by immunofluorescence. Supported by greater fluorescence in basal bodies than in adjacent axonemes in tracheal cells, analysis of basal apparatuses demonstrated both a proportionately greater ratio of tektin to tubulin (approximately 1:13) and two distinct solubility classes of tektins, consistent with tektins comprising the B-C junction of triplets in addition to the A-B junction as in doublets.
Department of Physiology, Boston University School of Medicine, Massachusetts 02118, USA. rstephen@bu.edu
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Stephens
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Lemieux
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification; D049872:Bivalvia; D002923:Cilia / Q000737:chemistry*; D015151:Immunoblotting; D008297:Male; D008856:Microscopy, Fluorescence; D008868:Microtubule Proteins / Q000032:analysis* / Q000276:immunology; D008974:Mollusca; D011817:Rabbits; D013082:Sperm Tail / Q000737:chemistry*
D000906:Antibodies; D008868:Microtubule Proteins