Transient electric birefringence study of intermediate filament formation from vimentin and glial fibrillary acidic protein.
Mg2+-induced polymerization of type III intermediate filament proteins vimentin and glial fibrillary acidic protein was studied by transient electric birefringence. In the absence of MgCl2 we found a net permanent dipole moment, approximately 45-nm-long dimers for vimentin, approximately 65-nm-long tetramers, hexamers, and possibly octamers for both proteins, and 100-nm aggregates for glial fibrillary acidic protein. Controlled oligomerization occurred after the addition of MgCl2. Although the solutions contained (small) aggregates of different sizes, more or less discrete steps in polymer formation were observed, and it was possible to discriminate between an increase in width and length. At the first stage of polymerization (in 0.3 mM MgCl2 for vimentin and 0.2 mM MgCl2 for glial fibrillary acidic protein), the permanent dipole moment disappeared without a change in length of the particles. At higher MgCl2 concentrations, structures of approximately 100 nm were formed, which strongly tended to laterally assemble into full-width intermediate filament structures consisting of about 32 monomers. This contrasts with previous models where first full-width (approximately 10-nm) aggregates are formed, which then increase in length. Subsequently, two discrete elongation steps of 35 nm are observed that increase the length to 135 and 170 nm, respectively. Possible structural models are suggested for the polymerization.
Kooijman
M
M
Department of Physics and Astronomy and Institute for Molecular Biological Sciences, Vrije Universiteit, De Boelelaan 1081, 1081 HV Amsterdam, The Netherlands.
Bloemendal
M
M
Traub
P
P
van Grondelle
R
R
van Amerongen
H
H
D000818:Animals; D001704:Biopolymers; D001718:Birefringence; D002417:Cattle; D004560:Electricity; D005904:Glial Fibrillary Acidic Protein / Q000737:chemistry*; D015636:Magnesium Chloride / Q000737:chemistry; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000737:chemistry*
D001704:Biopolymers; D005904:Glial Fibrillary Acidic Protein; D014746:Vimentin; D015636:Magnesium Chloride