Characterization of two distinct monoclonal antibodies to paired helical filaments: further evidence for fetal-type phosphorylation of the tau in paired helical filaments.
Two monoclonal antibodies C5 and M4 raised against Sarkosyl-insoluble paired helical filaments (PHF) specifically labeled fetal tau, but hardly labeled normal adult tau. C5 immunoreactivity was eliminated by alkaline phosphatase treatment at 37 degrees C, whereas M4 reactivity could be removed only by the treatment at 67 degrees C. Epitope analysis showed that C5 and M4 recognition sites are in residues 386-406 and 198-250, respectively, according to the numbering of the longest human tau isoform. Thus, the phosphorylation sites are located in the amino- and carboxyl-terminal portions of the microtubule-binding region. These two well-characterized monoclonals should be valuable in the identification of a protein kinase(s) that converts normal tau into PHF-tau.
Hasegawa
M
M
Department of Neuropathology, Faculty of Medicine, University of Tokyo, Japan.
Watanabe
A
A
Takio
K
K
Suzuki
M
M
Arai
T
T
Titani
K
K
Ihara
Y
Y
D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology; D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D002540:Cerebral Cortex / Q000378:metabolism / Q000473:pathology; D003902:Detergents; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005333:Fetus; D006801:Humans; D015151:Immunoblotting; D007382:Intermediate Filaments / Q000378:metabolism* / Q000648:ultrastructure; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D012521:Sarcosine / Q000031:analogs & derivatives; D017386:Sequence Homology, Amino Acid; D016875:tau Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D003902:Detergents; D000939:Epitopes; D016875:tau Proteins; C025231:sarkosyl; D011494:Protein Kinases; D012521:Sarcosine