Acceleration of bovine neurofilament L assembly by deprivation of acidic tail domain.
Functions of the tail region of neurofilament L have, to date, not been clearly elucidated. Bovine neurofilament L was cleaved into tail-less neurofilament L (50 kDa) and a tail fragment (19 kDa), by thrombin. Tail-less neurofilament L was deficit of the highly acidic domain of the tail region (approximately 77% of the entire region). Assembly of tail-less neurofilament L. was observed to be accelerated by both fluorometric and centrifugal measurements, compared with intact neurofilament L. The critical concentration of tail-less neurofilament L, which constitutes the constant unassembled pool, was approximately 0.25-times lower than that of neurofilament L. Under physiological conditions, tail-less neurofilament L formed a ribbon-like structure, whereas tail-less neurofilament L could form 10-nm filaments in an extremely low ionic-strength buffer in the presence of 1 mM MgCl2. An affinity-purified antibody directed against the tail fragment also accelerated neurofilament L assembly. The tail fragment neither coassembled with neurofilament L nor affect neurofilament L assembly. The acidic domain of the tail region may regulate neurofilament assembly and may be involved in 10-nm filament formation under physiological conditions.
Nakamura
Y
Y
Department of Neuropsychiatry, Osaka University Medical School, Fukushima-ku, Japan.
Takeda
M
M
Aimoto
S
S
Hariguchi
S
S
Kitajima
S
S
Nishimura
T
T
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016900:Neurofilament Proteins / Q000737:chemistry* / Q000276:immunology / Q000502:physiology; D009994:Osmolar Concentration
D016900:Neurofilament Proteins; C044882:neurofilament protein L; D008274:Magnesium