Quantitative analysis of tubulin and microtubule compartments in isolated rat hepatocytes.
A combined morphometric and biochemical approach has been used to identify and quantitate microtubules and tubulin in isolated hepatocytes. The total soluble pool of microtubule protein was estimated by specific high affinity binding to radiolabeled colchicine. Scatchard analysis of the data identified two populations of binding sites: high affinity-low capacity sites resembling tubulin and low affinity-high capacity sites believed to represent nonspecific colchicine-binding sites. Data from these studies indicate that tubulin represents 1% of the soluble protein of the cell, that 9.0 X 10(-14) dimers of tubulin are present per microgram soluble hepatocyte protein, and that the average hepatocyte contains 3.1 X 10(7) tubulin dimers. Our calculations suggest that this amount of tubulin would form a microtubule 1.9 cm in length if totally assembled. However, stereological measurements indicate that the actual length of microtubules in the cytosolic compartment of the average hepatocyte is only 0.28 cm. Thus, these experiments suggest that only 15% of the available tubulin in hepatocytes of postabsorptive rats is assembled in the form of microtubules.
D000818:Animals; D001665:Binding Sites; D002452:Cell Count; D003078:Colchicine / Q000378:metabolism; D003080:Cold Temperature; D006023:Glycoproteins / Q000032:analysis*; D008099:Liver / Q000032:analysis / Q000166:cytology* / Q000378:metabolism; D008870:Microtubules / Q000032:analysis*; D011034:Podophyllotoxin / Q000494:pharmacology; D051381:Rats; D014404:Tubulin / Q000032:analysis* / Q000378:metabolism; D014747:Vinblastine / Q000494:pharmacology
D006023:Glycoproteins; D014404:Tubulin; D014747:Vinblastine; D011034:Podophyllotoxin; D003078:Colchicine