Analysis of tubulin proteins and peptides in neuronal and non-neuronal tissues using immobilized pH gradients.
The accurate identification of the individual protein products of multi-gene families is essential to the interpretation of data from a wide range of experimental approaches including molecular biology, protein chemistry, and cell biology. We have adapted immobilized pH gradient isoelectric focusing to provide high resolution of tubulin proteins. Here we use these techniques to investigate the heterogeneity of tubulin in several neuronal and non-neuronal tissues to provide an accurate evaluation of isotubulin composition. Of the ten sources examined, the greatest number of isotubulins was found in whole adult brain. Tubulin isolated from either neonate human or rat brain consists predominantly of the more basic alpha and beta isotubulins found in adult brain. Cerebrum, cerebellum, medulla and caudate nucleus all contain the same large number of isotubulins as in whole brain, but in varying proportions. Liver, kidney and spleen isotubulin populations are all similar to each other and consist of a simpler distribution than any neuronal tissue examined. The majority of the tubulin protein in these non-neuronal tissues is composed of only the most basic alpha tubulins and intermediately-charged beta tubulins. No isotubulins were identified that were unique to these three non-neuronal tissues. Tubulin from neuroblastoma cells has an isotubulin distribution grossly similar to non-neuronal sources but additionally contains two basic beta isotubulins found in adult brain that are absent from non-neuronal tissues.
Field
D J
DJ
Department of Medical Genetics, University of Toronto, Ontario, Canada.
Lee
J C
JC
D000818:Animals; D000831:Animals, Newborn; D001923:Brain Chemistry; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D014644:Genetic Variation; D007525:Isoelectric Focusing / Q000379:methods; D009474:Neurons / Q000032:analysis; D009928:Organ Specificity; D010446:Peptide Fragments / Q000032:analysis*; D010449:Peptide Mapping; D051381:Rats; D012756:Sheep; D014404:Tubulin / Q000032:analysis* / Q000235:genetics
D010446:Peptide Fragments; D014404:Tubulin