Characterization of heterogeneous solutions using laser light scattering: study of the tubulin system

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Abstract
Laser light scattering was applied to a systematic study of a heterogeneous solution of [porcine brain] tubulin at low temperature conditions under which tubulin assembly into microtubules does not take place. Methods of analyzing laser light scattering results obtained from solutions containing multiple components are discussed. Data analysis techniques are described and their application to the determination of diffusion constants from experimental data is extensively illustrated. Multiple components were found under the conditions that the tubulin was studied. One component having D[particle self diffusion constant]20,w = 4.41 .times. 10-7 cm2/s (.sigma. = 0.54 .times. 10-7 cm2/s) which has the expected value for tubulin dimer was identified. Also found were 2 components which are significantly larger than tubulin. One large component has D20,w .apprx. 0.55 .times. 10-7 cm2/s and is present in all samples at 4.degree. C even after centrifugation to remove components greater than 10 S. Another large component having 3.2 .times. 10-7 cm2/s .gtoreq. D20,w .gtorsim. 1.5 .times. 10-7 cm2/s sedimented with 10 S .ltoreq. s < 20 S.

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