On the Identification of α‐ and β‐Tubulin Subunits

Abstract

Confusion appears to have arisen in the literature regarding the designation of .alpha.- and .beta.-tubulin in polyacrylamide gels. The presence or absence of 8 M urea in sodium dodecyl sulfate (SDS) polyacrylamide gels leads to different patterns for unalkylated tubulin subunits (and other proteins), making difficult the designation of the .alpha. and .beta. subunits by original definition using electrophoretic mobility in the MW dimension. The specific biochemical property of posttranslational tyrosylation of the .alpha. subunit in bullfrog dorsal root ganglion was used to identify further this subunit. Under all conditions tested, the .beta. subunit was more acidic than the .alpha. subunit, with isoelectric point differences that agree with theoretical and published values. If the tubulin subunits are reduced and alkylated, the .beta. subunit migrates more rapidly in SDS polyacrylamide gels, with or without urea present. Unalkylated tubulin subunits can comigrate or even reverse their relative mobility if 8 M urea-SDS polyacrylamide gels are used for subunit separation. Earlier reports are confirmed that the post-translational tyrosylation of protein appears exclusively restricted to .alpha.-tubulin and can be demonstrated in an in vivo situation. Only the .alpha.2 subunit of tubulin is tyrosylated.