Specific glycosaminoglycan chain length and sulfation patterns are required for cell uptake of tau versus -synuclein and -amyloid aggregates

Barbara E. Stopschinski, Brandon B. Holmes, Gregory M. Miller, Victor A. Manon, Jaime Vaquer-Alicea, William L. Prueitt, Linda C. Hsieh-Wilson, Marc I. Diamond

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Transcellular propagation of protein aggregate “seeds” has been proposed to mediate the progression of neurodegenerative diseases in tauopathies and -synucleinopathies. We previously reported that tau and -synuclein aggregates bind heparan sulfate proteoglycans (HSPGs) on the cell surface, promoting cellular uptake and intracellular seeding. However, the specificity and binding mode of these protein aggregates to HSPGs remain unknown. Here, we measured direct interaction with modified heparins to determine the size and sulfation requirements for tau, -synuclein, and -amyloid (A) aggregate binding to glycosaminoglycans (GAGs). Varying the GAG length and sulfation patterns, we next conducted competition studies with heparin derivatives in cell-based assays. Tau aggregates required a precise GAG architecture with defined sulfate moieties in the N- and 6-O-positions, whereas the binding of -synuclein and A aggregates was less stringent. To determine the genes required for aggregate uptake, we used CRISPR/Cas9 to individually knock out the major genes of the HSPG synthesis pathway in HEK293T cells. Knockouts of the extension enzymes exostosin 1 (EXT1), exostosin 2 (EXT2), and exostosin-like 3 (EXTL3), as well as N-sulfotransferase (NDST1) or 6-O-sulfotransferase (HS6ST2) significantly reduced tau uptake, consistent with our biochemical findings, and knockouts of EXT1, EXT2, EXTL3, or NDST1, but not HS6ST2 reduced -synuclein uptake. In summary, tau aggregates display specific interactions with HSPGs that depend on GAG length and sulfate moiety position, whereas -synuclein and A aggregates exhibit more flexible interactions with HSPGs. These principles may inform the development of mechanism-based therapies to block transcel-lular propagation of amyloid protein–based pathologies.

Original languageEnglish (US)
Pages (from-to)10826-10840
Number of pages15
JournalJournal of Biological Chemistry
Volume293
Issue number27
DOIs
StatePublished - Jan 1 2018

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Synucleins
Heparan Sulfate Proteoglycans
Glycosaminoglycans
Chain length
Amyloid
Sulfotransferases
Sulfates
Heparin
Clustered Regularly Interspaced Short Palindromic Repeats
Tauopathies
Gene Knockout Techniques
Genes
Neurodegenerative diseases
Neurodegenerative Diseases
Seeds
Carrier Proteins
Pathology
Seed
Assays
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Specific glycosaminoglycan chain length and sulfation patterns are required for cell uptake of tau versus -synuclein and -amyloid aggregates. / Stopschinski, Barbara E.; Holmes, Brandon B.; Miller, Gregory M.; Manon, Victor A.; Vaquer-Alicea, Jaime; Prueitt, William L.; Hsieh-Wilson, Linda C.; Diamond, Marc I.

In: Journal of Biological Chemistry, Vol. 293, No. 27, 01.01.2018, p. 10826-10840.

Research output: Contribution to journalArticle

Stopschinski, Barbara E. ; Holmes, Brandon B. ; Miller, Gregory M. ; Manon, Victor A. ; Vaquer-Alicea, Jaime ; Prueitt, William L. ; Hsieh-Wilson, Linda C. ; Diamond, Marc I. / Specific glycosaminoglycan chain length and sulfation patterns are required for cell uptake of tau versus -synuclein and -amyloid aggregates. In: Journal of Biological Chemistry. 2018 ; Vol. 293, No. 27. pp. 10826-10840.
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AU - Manon, Victor A.

AU - Vaquer-Alicea, Jaime

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AU - Diamond, Marc I.

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