Different reactivities of brain and erythrocyte tubulins toward a sulfhydryl group-directed reagent that inhibits microtubule assembly.
There is considerable evidence that tubulin exists in multiple isotypes, differing in amino acid sequence and tissue distribution. Little is known, however, about the functional significance of these isotypes. Chicken erythrocyte beta-tubulin has been shown by peptide mapping to differ significantly from chicken brain beta-tubulin (Murphy, D. B., and Wallis, K. T. (1983) J. Biol. Chem. 258, 7870-7875). We now find that when the two tubulins, in their native states, are incubated with N,N'-ethylenebis(iodoacetamide) (EBI), a bifunctional sulfhydryl-directed reagent, microtubule assembly by brain tubulin is much more sensitive to inhibition by EBI than is erythrocyte tubulin assembly. The resistance of erythrocyte microtubule assembly to inhibition by EBI is correlated with a low reactivity of erythrocyte tubulin with [14C]EBI. This difference is most marked in the beta subunit which reacts 15 and 17% as well, respectively, with [14C]EBI as do the beta 1 and beta 2 subunits of brain tubulin. Also, erythrocyte beta reacts about 33% as well as does brain beta with iodo[14C]acetamide. These results suggest that a reactive sulfhydryl group, whose oxidation prevents microtubule assembly, is present in brain tubulin but absent or inaccessible in erythrocyte tubulin. Since purified erythrocyte tubulin self-aggregates much more readily than does brain tubulin, it is conceivable that erythrocyte and brain tubulin may differ in that the latter may have its assembly subject to a complex regulation, while erythrocyte tubulin assembly may be regulated by a simpler mechanism.
D000818:Animals; D001923:Brain Chemistry; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000032:analysis*; D005029:Ethylenediamines / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D007460:Iodoacetamide / Q000494:pharmacology; D007700:Kinetics; D008854:Microscopy, Electron; D008870:Microtubules / Q000187:drug effects*; D011108:Polymers / Q000378:metabolism; D013329:Structure-Activity Relationship; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D014404:Tubulin / Q000378:metabolism*; D014747:Vinblastine / Q000494:pharmacology
D005029:Ethylenediamines; D011108:Polymers; D013439:Sulfhydryl Reagents; D014404:Tubulin; D014747:Vinblastine; C032156:N,N'-ethylenebis(iodoacetamide); D006160:Guanosine Triphosphate; D007460:Iodoacetamide