Tissue transglutaminase modulates alpha-synuclein oligomerization

Gezina M.J. Segers-Nolten, Micha M.M. Wilhelmus, G.J. Veldhuis, G. Veldhuis, Bart van Rooijen, Benjamin Drukarch, Vinod Subramaniam

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

We have studied the interaction of the enzyme tissue transglutaminase (tTG), catalyzing cross-link formation between protein-bound glutamine residues and primary amines, with Parkinson's disease-associated α-synuclein protein variants at physiologically relevant concentrations. We have, for the first time, determined binding affinities of tTG for wild-type and mutant α-synucleins using surface plasmon resonance approaches, revealing high-affinity nanomolar equilibrium dissociation constants. Nanomolar tTG concentrations were sufficient for complete inhibition of fibrillization by effective α-synuclein cross-linking, resulting predominantly in intramolecularly cross-linked monomers accompanied by an oligomeric fraction. Since oligomeric species have a pathophysiological relevance we further investigated the properties of the tTG/α-synuclein oligomers. Atomic force microscopy revealed morphologically similar structures for oligomers from all α-synuclein variants; the extent of oligomer formation was found to correlate with tTG concentration. Unlike normal α-synuclein oligomers the resultant structures were extremely stable and resistant to GdnHCl and SDS. In contrast to normal β-sheet-containing oligomers, the tTG/α-synuclein oligomers appear to be unstructured and are unable to disrupt phospholipid vesicles. These data suggest that tTG binds equally effective to wild-type and disease mutant α-synuclein variants. We propose that tTG cross-linking imposes structural constraints on α-synuclein, preventing the assembly of structured oligomers required for disruption of membranes and for progression into fibrils. In general, cross-linking of amyloid forming proteins by tTG may prevent the progression into pathogenic species.
Original languageUndefined
Pages (from-to)1395-1402
Number of pages8
JournalProtein science
Volume17
Issue number8
DOIs
Publication statusPublished - 2008

Keywords

  • Parkinson's Disease
  • oligomer
  • Surface plasmon resonance
  • tissue transglutaminase
  • α-Synuclein
  • METIS-248984
  • IR-72349
  • Cross-linking
  • Atomic Force Microscopy

Cite this

Segers-Nolten, G. M. J., Wilhelmus, M. M. M., Veldhuis, G. J., Veldhuis, G., van Rooijen, B., Drukarch, B., & Subramaniam, V. (2008). Tissue transglutaminase modulates alpha-synuclein oligomerization. Protein science, 17(8), 1395-1402. https://doi.org/10.1110/ps.036103.108
Segers-Nolten, Gezina M.J. ; Wilhelmus, Micha M.M. ; Veldhuis, G.J. ; Veldhuis, G. ; van Rooijen, Bart ; Drukarch, Benjamin ; Subramaniam, Vinod. / Tissue transglutaminase modulates alpha-synuclein oligomerization. In: Protein science. 2008 ; Vol. 17, No. 8. pp. 1395-1402.
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abstract = "We have studied the interaction of the enzyme tissue transglutaminase (tTG), catalyzing cross-link formation between protein-bound glutamine residues and primary amines, with Parkinson's disease-associated α-synuclein protein variants at physiologically relevant concentrations. We have, for the first time, determined binding affinities of tTG for wild-type and mutant α-synucleins using surface plasmon resonance approaches, revealing high-affinity nanomolar equilibrium dissociation constants. Nanomolar tTG concentrations were sufficient for complete inhibition of fibrillization by effective α-synuclein cross-linking, resulting predominantly in intramolecularly cross-linked monomers accompanied by an oligomeric fraction. Since oligomeric species have a pathophysiological relevance we further investigated the properties of the tTG/α-synuclein oligomers. Atomic force microscopy revealed morphologically similar structures for oligomers from all α-synuclein variants; the extent of oligomer formation was found to correlate with tTG concentration. Unlike normal α-synuclein oligomers the resultant structures were extremely stable and resistant to GdnHCl and SDS. In contrast to normal β-sheet-containing oligomers, the tTG/α-synuclein oligomers appear to be unstructured and are unable to disrupt phospholipid vesicles. These data suggest that tTG binds equally effective to wild-type and disease mutant α-synuclein variants. We propose that tTG cross-linking imposes structural constraints on α-synuclein, preventing the assembly of structured oligomers required for disruption of membranes and for progression into fibrils. In general, cross-linking of amyloid forming proteins by tTG may prevent the progression into pathogenic species.",
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Segers-Nolten, GMJ, Wilhelmus, MMM, Veldhuis, GJ, Veldhuis, G, van Rooijen, B, Drukarch, B & Subramaniam, V 2008, 'Tissue transglutaminase modulates alpha-synuclein oligomerization' Protein science, vol. 17, no. 8, pp. 1395-1402. https://doi.org/10.1110/ps.036103.108

Tissue transglutaminase modulates alpha-synuclein oligomerization. / Segers-Nolten, Gezina M.J.; Wilhelmus, Micha M.M.; Veldhuis, G.J.; Veldhuis, G.; van Rooijen, Bart; Drukarch, Benjamin; Subramaniam, Vinod.

In: Protein science, Vol. 17, No. 8, 2008, p. 1395-1402.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Tissue transglutaminase modulates alpha-synuclein oligomerization

AU - Segers-Nolten, Gezina M.J.

AU - Wilhelmus, Micha M.M.

AU - Veldhuis, G.J.

AU - Veldhuis, G.

AU - van Rooijen, Bart

AU - Drukarch, Benjamin

AU - Subramaniam, Vinod

PY - 2008

Y1 - 2008

N2 - We have studied the interaction of the enzyme tissue transglutaminase (tTG), catalyzing cross-link formation between protein-bound glutamine residues and primary amines, with Parkinson's disease-associated α-synuclein protein variants at physiologically relevant concentrations. We have, for the first time, determined binding affinities of tTG for wild-type and mutant α-synucleins using surface plasmon resonance approaches, revealing high-affinity nanomolar equilibrium dissociation constants. Nanomolar tTG concentrations were sufficient for complete inhibition of fibrillization by effective α-synuclein cross-linking, resulting predominantly in intramolecularly cross-linked monomers accompanied by an oligomeric fraction. Since oligomeric species have a pathophysiological relevance we further investigated the properties of the tTG/α-synuclein oligomers. Atomic force microscopy revealed morphologically similar structures for oligomers from all α-synuclein variants; the extent of oligomer formation was found to correlate with tTG concentration. Unlike normal α-synuclein oligomers the resultant structures were extremely stable and resistant to GdnHCl and SDS. In contrast to normal β-sheet-containing oligomers, the tTG/α-synuclein oligomers appear to be unstructured and are unable to disrupt phospholipid vesicles. These data suggest that tTG binds equally effective to wild-type and disease mutant α-synuclein variants. We propose that tTG cross-linking imposes structural constraints on α-synuclein, preventing the assembly of structured oligomers required for disruption of membranes and for progression into fibrils. In general, cross-linking of amyloid forming proteins by tTG may prevent the progression into pathogenic species.

AB - We have studied the interaction of the enzyme tissue transglutaminase (tTG), catalyzing cross-link formation between protein-bound glutamine residues and primary amines, with Parkinson's disease-associated α-synuclein protein variants at physiologically relevant concentrations. We have, for the first time, determined binding affinities of tTG for wild-type and mutant α-synucleins using surface plasmon resonance approaches, revealing high-affinity nanomolar equilibrium dissociation constants. Nanomolar tTG concentrations were sufficient for complete inhibition of fibrillization by effective α-synuclein cross-linking, resulting predominantly in intramolecularly cross-linked monomers accompanied by an oligomeric fraction. Since oligomeric species have a pathophysiological relevance we further investigated the properties of the tTG/α-synuclein oligomers. Atomic force microscopy revealed morphologically similar structures for oligomers from all α-synuclein variants; the extent of oligomer formation was found to correlate with tTG concentration. Unlike normal α-synuclein oligomers the resultant structures were extremely stable and resistant to GdnHCl and SDS. In contrast to normal β-sheet-containing oligomers, the tTG/α-synuclein oligomers appear to be unstructured and are unable to disrupt phospholipid vesicles. These data suggest that tTG binds equally effective to wild-type and disease mutant α-synuclein variants. We propose that tTG cross-linking imposes structural constraints on α-synuclein, preventing the assembly of structured oligomers required for disruption of membranes and for progression into fibrils. In general, cross-linking of amyloid forming proteins by tTG may prevent the progression into pathogenic species.

KW - Parkinson's Disease

KW - oligomer

KW - Surface plasmon resonance

KW - tissue transglutaminase

KW - α-Synuclein

KW - METIS-248984

KW - IR-72349

KW - Cross-linking

KW - Atomic Force Microscopy

U2 - 10.1110/ps.036103.108

DO - 10.1110/ps.036103.108

M3 - Article

VL - 17

SP - 1395

EP - 1402

JO - Protein science

JF - Protein science

SN - 0961-8368

IS - 8

ER -

Segers-Nolten GMJ, Wilhelmus MMM, Veldhuis GJ, Veldhuis G, van Rooijen B, Drukarch B et al. Tissue transglutaminase modulates alpha-synuclein oligomerization. Protein science. 2008;17(8):1395-1402. https://doi.org/10.1110/ps.036103.108