Oxidation of cysteine-322 in the repeat domain of microtubule-associated protein tau controls the in vitro assembly of paired helical filaments.
One of the hallmarks of Alzheimer disease is the pathological aggregation of tau protein into paired helical filaments (PHFs) and neurofibrillary tangles. Here we describe the in vitro assembly of recombinant tau protein and constructs derived from it into PHFs. Though whole tau assembled poorly, constructs containing three internal repeats (corresponding to the fetal tau isoform) formed PHFs reproducibly. This ability depended on intermolecular disulfide bridges formed by the single Cys-322. Blocking the SH group, mutating Cys for Ala, or keeping tau in a reducing environment all inhibited assembly. With constructs derived from four-repeat tau (having the additional repeat no. 2 and a second Cys-291), PHF assembly was blocked because Cys-291 and Cys-322 interact within the molecule. PHF assembly was enabled again by mutating Cys-291 for Ala. The synthetic PHFs bound the dye thioflavin S used in Alzheimer disease diagnostics. The data imply that the redox potential in the neuron is crucial for PHF assembly, independently or in addition to pathological phosphorylation reactions.
Schweers
O
O
Max-Planck Unit for Structural Molecular Biology, c/o Deutsches Elektronen Synchroton, Hamburg, Germany.
Mandelkow
E M
EM
Biernat
J
J
Mandelkow
E
E
D000544:Alzheimer Disease / Q000378:metabolism* / Q000473:pathology; D052160:Benzothiazoles; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D016874:Neurofibrillary Tangles / Q000378:metabolism; D010084:Oxidation-Reduction; D010766:Phosphorylation; D011487:Protein Conformation; D013439:Sulfhydryl Reagents / Q000737:chemistry; D013844:Thiazoles / Q000737:chemistry; D016875:tau Proteins / Q000737:chemistry / Q000378:metabolism*
D052160:Benzothiazoles; D005456:Fluorescent Dyes; D013439:Sulfhydryl Reagents; D013844:Thiazoles; D016875:tau Proteins; C009462:thioflavin T; D003545:Cysteine