Palmitoylation of tubulin.
Tubulin is a very water soluble protein, yet a significant portion is firmly associated with cell membranes. Because recent work has shown that palmitoylation is a dynamic process that can alter the targeting of proteins to membranes, we tested whether or not tubulin could be palmitoylated to account for its membrane location. Tubulin acylation was measured by incorporation of [3H]palmitate into PC12 cells in culture. We found palmitoylated tubulin in both cell pellet and cytosol with a higher concentration in the former. EGF-stimulated PC12 cells incorporated the same amount of palmitate per unit protein but the proportion in the membrane fraction was enhanced. More palmitate of the pellet was found in alpha than beta tubulin; EGF stimulation primarily increased palmitate in beta tubulin. In addition we found that palmitic acid was present both as thioesters and as oxyesters. We suggest that palmitoylation may contribute to the membrane localization of tubulin and can be regulated by growth factors.
Zambito
A M
AM
Laboratory of Biochemical Pharmacology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892, USA.
Wolff
J
J
D000818:Animals; D004396:Coloring Agents; D004591:Electrophoresis, Polyacrylamide Gel; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016716:PC12 Cells; D010169:Palmitic Acids / Q000378:metabolism*; D051381:Rats; D012394:Rosaniline Dyes; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014316:Tritium; D014404:Tubulin / Q000187:drug effects / Q000378:metabolism*
D004396:Coloring Agents; D010169:Palmitic Acids; D012394:Rosaniline Dyes; D013438:Sulfhydryl Compounds; D014404:Tubulin; D014316:Tritium; D004815:Epidermal Growth Factor; C048139:Coomassie blue