Calpain‐Induced Proteolysis of Normal Human Tau and Tau Associated with Paired Helical Filaments
Open accessAbstract
The major components of neurofibrillary tangles (NFT) in Alzheimer's disease are bundles of paired helical filaments (PHF) which are primarily composed of highly phosphorylated tau proteins (PHF‐tau). To further understand the mechanism of PHF accumulation in NFT, we examined the calpain‐induced proteolysis of highly purified and primarily non‐aggregated PHF and normal tau proteins with various contents of phosphate isolated from either fetal (F‐tau) or adult human brain (N‐tau). The extent of proteolysis was determined by decreases in tau immunoreactivity using Western‐blot analysis and a panel of site‐specific tau antibodies (Alz 50, Tau‐2, Tau 14, Tau‐1, AT8, E‐11, AH‐1 and PHF‐1). We found that full‐size polypeptides of N‐tau and F‐tau were similarly and rapidly proteolyzed in vitro by calpain (calpain II, 3.3 units/mg protein) during a 10‐min incubation at 30°C, and that their half lives (t1/2) were 1.5 min and 1.8 min, respectively, Analysis of immunoblots suggests that full‐length polypeptides of tau are first degraded into large fragments similar in size to that generated endogenously, then into smaller fragments. Since both endogenous and in‐vitro‐generated tau fragments retained N‐terminal epitopes, the results suggest that most of the calpain‐sensitive sites may be located in the C‐terminal half of the tau molecule. In contrast, PHF were extremely resistant to degradation and only a fivefold higher concentration of calpain (16.7 units/mg protein) induced partial proteolysis of PHF. A major calpain‐generated fragment was a 45‐kDa polypeptide derived from the C‐terminal region of PHF‐tau, which forms a core of filaments. The results suggest that the inaccessibility of potential calpain‐digestion sites in the filament core could contribute to the resistance of PHF to calpain and subsequently lead to the accumulation of PHF in Alzheimer's disease. The results also suggest that hyperphosphorylation of tau may be mariginally involved in the resistance of PHF to degradation by calpain. Ultrastructural examination revealed that, in contrast to previous studies with trypsin, calpain did not alter the morphologic appearance of filaments; after incubation with calpain, the majority of PHF remained short and disperse and the number of PHF aggregated into NFT‐like clusters was not significantly increased. The results suggest that the role of calpain in promoting the aggregation and clustering of filaments is limited.
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Calpain-1 cleaves tau at Lys44, Arg230, Arg242, Gly323, and Gly326.
“Calpain-1 cleaves tau at Lys44, Arg230, Arg242, Gly323, and Gly326”
From Molecular and cellular mechanisms underlying the pathogenesis of Alzheimer’s disease (2020) · cites this paper as [203]
Calpain-2 cleaves tau at Arg230.
“while calpain-2 cleaves tau at Arg230 [202–206].”
From Molecular and cellular mechanisms underlying the pathogenesis of Alzheimer’s disease (2020) · cites this paper as [203]
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- Authors
- Yang, L., & Ksiezak‐Reding, H.
- Affiliations
- Albert Einstein College of Medicine (US)
- Venue
- European Journal of Biochemistry
- Publisher
- Wiley
- Published
- 1995-10
- Volume
- 233
- Issue
- 1
- Pages
- 9-17
- ISSN
- 0014-2956 (ISSN-L)
- 1432-1033
- OpenAlex
- W2021017365
- Cited by
- 113
- References
- 57
- Citations by year
- 2025: 2 · 2023: 4 · 2022: 6 · 2021: 6 · 2020: 5 · 2019: 2 · 2018: 4 · 2017: 3 · 2016: 1 · 2015: 5 · 2014: 2 · 2013: 5 · 2012: 5
- FWCI
- 2.87
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- European Journal of Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01ns030027-03)
- National Institute of Neurological Disorders and Stroke (NS30027)
- Topics
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Calpain Protease Function and Regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › S100 Proteins and Annexins
- Keywords
- calpain, tau protein, paired helical filaments, neurofibrillary tangles, Alzheimer's disease, proteolysis, tau hyperphosphorylation, C-terminal cleavage, tau phosphorylation sites, trypsin digestion, site-specific antibodies
- MeSH terms
- Adult, Aged, Alzheimer Disease, Binding Sites, Brain Chemistry, Calpain, Female, Fetus, Humans, Male, Microscopy, Electron, Peptide Fragments, Phosphorylation, Neurofibrillary Tangles, tau Proteins
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