title: Electrophoretic resolution of tubulin and tektin subunits by differential interaction with long-chain alkyl sulfates.
abstract: Tubulin dimers, formed from globular alpha and beta subunits, and the tektins, three equimolar alpha-helical proteins that form filaments, mutually associate to form the junctional regions of doublet and triplet microtubules. When evaluated by SDS-PAGE, the apparent molecular weights of these proteins can deviate substantially from their sequence molecular weights in a manner sensitive to both the source of SDS and the species of origin. The electrophoretic mobility of sperm tail flagellar tubulins and tektins from an echinoderm and a mollusc were studied systematically using detergent-free stacking and resolving gels with a running buffer containing pure sodium dodecyl sulfate augmented with fixed amounts of C10, C14, C16, or C18 alkyl sulfates. Although having no systematic effect on molecular weight standards, the presence of alkyl sulfates of increasing chain length progressively exaggerated the separation of tubulin subunits, similarly facilitated the separation of two normally comigrating tektins, yet minimally influenced the relative migration of adequately separated tektins. This phenomenon is most likely due to preferential binding of longer chain alkyl sulfates by specific hydrophobic regions of these otherwise similar proteins. The use of binary mixtures of pure alkyl sulfates, required in the running buffer alone, may prove useful for reproducibly separating other proteins that characteristically bind SDS anomalously.
authors.affiliation: Department of Physiology, Boston University School of Medicine, 715 Albany Street, Boston, Massachusetts, 02118, USA.
authors.forename: R E
authors.initials: RE
authors.lastname: Stephens
mesh_terms: D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D008868:Microtubule Proteins / Q000737:chemistry / Q000302:isolation & purification*; D014404:Tubulin / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D008868:Microtubule Proteins; D014404:Tubulin; C044103:tektins