Membrane-bound tubulin: Resistance to cathepsin D and susceptibility to thrombin.

In recent studies we found that cytoplasmic tubulin from brain was rapidly split by brain cathepsin D. Two pools could be established; the major portion was split at 18%/h, a minor portion at 2%/h, under our experimental circumstances. In the present work these experiments were extended to membrane-bound tubulin from brain. The membrane-bound form, in contrast to the cytoplasmic tubulin, was not degraded by cerebral cathepsin D under similar experimental conditions. This was not due to the presence of an inhibitory protein since added cytoplasmic tubulin was degraded. Several other protein components of membrane fractions (synaptosomal, mitochondrial) were degraded by cathepsin D, as measured on two-dimensional electropherograms. Thrombin degraded cytoplasmic tubulin, but the degradation products differed from those of cathepsin D degradation. Thrombin also hydrolyzed membrane-bound tubulin, but at a lower rate than the cytoplasmic form. Our results indicate great differences in the breakdown rate of a protein, which depend on its localization, in accord with the differences found in in vivo turnover rates.