TY - JOUR

T1 - Cascade autohydrolysis of Alzheimer's Aβ peptides

AU - Wolfram, Martin

AU - Tiwari, Manish K.

AU - Hassenkam, Tue

AU - Li, Ming

AU - Bjerrum, Morten J.

AU - Meldal, Morten

PY - 2023

Y1 - 2023

N2 - Protein/peptide self-assembly into amyloid structures associates with major neurodegenerative disorders such as Alzheimer's disease (AD). Soluble assemblies (oligomers) of the A beta peptide and their aggregates are perceived as neurotoxic species in AD. While screening for synthetic cleavage agents that could break down such aberrant assemblies through hydrolysis, we observed that the assemblies of A beta oligopeptides, containing the nucleation sequence A beta(14-24) (H(14)QKLVFFAEDV(24)), could act as cleavage agents by themselves. Autohydrolysis showed a common fragment fingerprint among various mutated A beta(14-24) oligopeptides, A beta(12-25)-Gly and A beta(1-28), and full-length A beta(1-40/42), under physiologically relevant conditions. Primary endoproteolytic autocleavage at the Gln(15)-Lys(16), Lys(16)-Leu(17) and Phe(19)-Phe(20) positions was followed by subsequent exopeptidase self-processing of the fragments. Control experiments with homologous d-amino acid enantiomers A beta(12-25)-Gly and A beta(16-25)-Gly showed the same autocleavage pattern under similar reaction conditions. The autohydrolytic cascade reaction (ACR) was resilient to a broad range of conditions (20-37 degrees C, 10-150 mu M peptide concentration at pH 7.0-7.8). Evidently, assemblies of the primary autocleavage fragments acted as structural/compositional templates (autocatalysts) for self-propagating autohydrolytic processing at the A beta(16-21) nucleation site, showing the potential for cross-catalytic seeding of the ACR in larger A beta isoforms (A beta(1-28) and A beta(1-40/42)). This result may shed new light on A beta behaviour in solution and might be useful in the development of intervention strategies to decompose or inhibit neurotoxic A beta assemblies in AD.

AB - Protein/peptide self-assembly into amyloid structures associates with major neurodegenerative disorders such as Alzheimer's disease (AD). Soluble assemblies (oligomers) of the A beta peptide and their aggregates are perceived as neurotoxic species in AD. While screening for synthetic cleavage agents that could break down such aberrant assemblies through hydrolysis, we observed that the assemblies of A beta oligopeptides, containing the nucleation sequence A beta(14-24) (H(14)QKLVFFAEDV(24)), could act as cleavage agents by themselves. Autohydrolysis showed a common fragment fingerprint among various mutated A beta(14-24) oligopeptides, A beta(12-25)-Gly and A beta(1-28), and full-length A beta(1-40/42), under physiologically relevant conditions. Primary endoproteolytic autocleavage at the Gln(15)-Lys(16), Lys(16)-Leu(17) and Phe(19)-Phe(20) positions was followed by subsequent exopeptidase self-processing of the fragments. Control experiments with homologous d-amino acid enantiomers A beta(12-25)-Gly and A beta(16-25)-Gly showed the same autocleavage pattern under similar reaction conditions. The autohydrolytic cascade reaction (ACR) was resilient to a broad range of conditions (20-37 degrees C, 10-150 mu M peptide concentration at pH 7.0-7.8). Evidently, assemblies of the primary autocleavage fragments acted as structural/compositional templates (autocatalysts) for self-propagating autohydrolytic processing at the A beta(16-21) nucleation site, showing the potential for cross-catalytic seeding of the ACR in larger A beta isoforms (A beta(1-28) and A beta(1-40/42)). This result may shed new light on A beta behaviour in solution and might be useful in the development of intervention strategies to decompose or inhibit neurotoxic A beta assemblies in AD.

KW - SECONDARY STRUCTURE

KW - COLD DENATURATION

KW - COMMON MECHANISM

KW - ALPHA-SYNUCLEIN

KW - PROTEIN

KW - HYDROLYSIS

KW - DISEASE

KW - OLIGOMERS

KW - NEUROTOXICITY

KW - FRAGMENTS

U2 - 10.1039/d2sc06668h

DO - 10.1039/d2sc06668h

M3 - Journal article

C2 - 37206405

VL - 14

SP - 4986

EP - 4996

JO - Chemical Science

JF - Chemical Science

SN - 2041-6520

IS - 19

ER -