High-yield purification of fetal tau preserving its structure and phosphorylation pattern.
The fetal type of tau phosphorylation always re-appears during pathogenesis of Alzheimer's disease and related tauopathies. The major obstacle in the study of the fetal tau phosphorylation has been the lack of a simple and reproducible purification method yielding fetal tau with high purity and unmodified phosphorylation pattern. We have developed a two-step, highly efficient purification procedure of perchloric acid-extracted fetal tau by immunoaffinity chromatography and trichloroacetic acid (TCA) precipitation. The method yielded tau with more than 90% purity. Most importantly, purified fetal tau exhibited unmodified phosphorylation pattern as confirmed by phosphorylation-dependent antibodies. In summary, this purification process preserves and protects unstable phosphoresidues from dephosphorylation and allows their detailed molecular analysis especially in the pathogenesis of Alzheimer's disease and related tauopathies.
Ivanovova
Natalia
N
Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava, Slovak Republic.
Handzusova
Martina
M
Hanes
Jozef
J
Kontsekova
Eva
E
Novak
Michal
M
D000544:Alzheimer Disease / Q000276:immunology; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D001923:Brain Chemistry / Q000276:immunology; D010472:Perchlorates / Q000737:chemistry; D010766:Phosphorylation; D051381:Rats; D014238:Trichloroacetic Acid / Q000737:chemistry; D016875:tau Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*
D000906:Antibodies; D010472:Perchlorates; D016875:tau Proteins; D014238:Trichloroacetic Acid