Discrete soluble forms of middle and high molecular weight neurofilament proteins in dilute aqueous buffers.

Neurofilament proteins purified from bovine spinal cord were characterized by sedimentation studies in aqueous buffers. In 10 mM Tris, pH 8, the middle molecular weight neurofilament protein (NF-M) has a sedimentation coefficient, S20,w, of 2.6 S. Sedimentation equilibrium data shows considerable nonideality; extrapolation to infinite dilution and correction for the primary charge effect yield a molecular weight of 1.09 X 10(5), indicative of a monomeric structure. When the ionic strength was increased, the sedimentation coefficient increased slightly, and the protein began to form larger aggregates. Reconstitution of short intermediate filaments was observed upon dialysis of denatured NF-M versus a reconstitution buffer. A circular dichroism spectrum of NF-M in 10 mM Tris was typical of alpha + beta proteins. High molecular weight neurofilament protein (NF-H) showed a considerable tendency to aggregate in 10 mM Tris, but a principal species with a sedimentation coefficient of 3.2 S was observed, and sedimentation equilibrium data also suggest a monomeric structure.