The 17-residue-long N terminus in huntingtin controls stepwise aggregation in solution and on membranes via different mechanisms
© 2018 by The American Society for Biochemistry and Molecular Biology, Inc..
Aggregation of huntingtin protein arising from expanded polyglutamine (polyQ) sequences in the exon-1 region of mutant huntingtin plays a central role in the pathogenesis of Huntington's disease. The huntingtin aggregation pathways are of therapeutic and diagnostic interest, but obtaining critical information from the physiologically relevant htt exon-1 (Httex1) protein has been challenging. Using biophysical techniques and an expression and purification protocol that generates clean, monomeric Httex1, we identified and mapped three distinct aggregation pathways: 1) unseeded in solution; 2) seeded in solution; and 3) membrane-mediated. In solution, aggregation proceeded in a highly stepwise manner, in which the individual domains (N terminus containing 17 amino acids (N17), polyQ, and proline-rich domain (PRD)) become ordered at very different rates. The aggregation was initiated by an early oligomer requiring a pathogenic, expanded Gln length and N17 α-helix formation. In the second phase, β-sheet forms in the polyQ. The slowest step is the final structural maturation of the PRD. This stepwise mechanism could be bypassed by seeding, which potently accelerated aggregation and was a prerequisite for prion-like spreading in vivo Remarkably, membranes could catalyze aggregation even more potently than seeds, in a process that caused significant membrane damage. The N17 governed membrane-mediated aggregation by anchoring Httex1 to the membrane, enhancing local concentration and promoting collision via two-dimensional diffusion. Considering its central roles in solution and in membrane-mediated aggregation, the N17 represents an attractive target for inhibiting multiple pathways. Our approach should help evaluate such inhibitors and identify diagnostic markers for the misfolded forms identified here.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2018 |
|---|---|
| Erschienen: |
2018 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:293 |
|---|---|
| Enthalten in: |
The Journal of biological chemistry - 293(2018), 7 vom: 16. Feb., Seite 2597-2605 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Pandey, Nitin K [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 05.02.2019 Date Revised 05.11.2023 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1074/jbc.M117.813667 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM279457707 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM279457707 | ||
| 003 | DE-627 | ||
| 005 | 20231225023016.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2018 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1074/jbc.M117.813667 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n0931.xml |
| 035 | |a (DE-627)NLM279457707 | ||
| 035 | |a (NLM)29282287 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Pandey, Nitin K |e verfasserin |4 aut | |
| 245 | 1 | 4 | |a The 17-residue-long N terminus in huntingtin controls stepwise aggregation in solution and on membranes via different mechanisms |
| 264 | 1 | |c 2018 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 05.02.2019 | ||
| 500 | |a Date Revised 05.11.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2018 by The American Society for Biochemistry and Molecular Biology, Inc. | ||
| 520 | |a Aggregation of huntingtin protein arising from expanded polyglutamine (polyQ) sequences in the exon-1 region of mutant huntingtin plays a central role in the pathogenesis of Huntington's disease. The huntingtin aggregation pathways are of therapeutic and diagnostic interest, but obtaining critical information from the physiologically relevant htt exon-1 (Httex1) protein has been challenging. Using biophysical techniques and an expression and purification protocol that generates clean, monomeric Httex1, we identified and mapped three distinct aggregation pathways: 1) unseeded in solution; 2) seeded in solution; and 3) membrane-mediated. In solution, aggregation proceeded in a highly stepwise manner, in which the individual domains (N terminus containing 17 amino acids (N17), polyQ, and proline-rich domain (PRD)) become ordered at very different rates. The aggregation was initiated by an early oligomer requiring a pathogenic, expanded Gln length and N17 α-helix formation. In the second phase, β-sheet forms in the polyQ. The slowest step is the final structural maturation of the PRD. This stepwise mechanism could be bypassed by seeding, which potently accelerated aggregation and was a prerequisite for prion-like spreading in vivo Remarkably, membranes could catalyze aggregation even more potently than seeds, in a process that caused significant membrane damage. The N17 governed membrane-mediated aggregation by anchoring Httex1 to the membrane, enhancing local concentration and promoting collision via two-dimensional diffusion. Considering its central roles in solution and in membrane-mediated aggregation, the N17 represents an attractive target for inhibiting multiple pathways. Our approach should help evaluate such inhibitors and identify diagnostic markers for the misfolded forms identified here | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Huntington disease | |
| 650 | 4 | |a amyloid | |
| 650 | 4 | |a electron paramagnetic resonance (EPR) | |
| 650 | 4 | |a huntingtin exon-1 | |
| 650 | 4 | |a polyglutamine | |
| 650 | 4 | |a protein aggregation | |
| 650 | 7 | |a HTT protein, human |2 NLM | |
| 650 | 7 | |a Huntingtin Protein |2 NLM | |
| 650 | 7 | |a Peptides |2 NLM | |
| 650 | 7 | |a Protein Aggregates |2 NLM | |
| 650 | 7 | |a polyglutamine |2 NLM | |
| 650 | 7 | |a 26700-71-0 |2 NLM | |
| 700 | 1 | |a Isas, J Mario |e verfasserin |4 aut | |
| 700 | 1 | |a Rawat, Anoop |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Rachel V |e verfasserin |4 aut | |
| 700 | 1 | |a Langen, Jennifer |e verfasserin |4 aut | |
| 700 | 1 | |a Pandey, Priyatama |e verfasserin |4 aut | |
| 700 | 1 | |a Langen, Ralf |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t The Journal of biological chemistry |d 1945 |g 293(2018), 7 vom: 16. Feb., Seite 2597-2605 |w (DE-627)NLM000004995 |x 1083-351X |7 nnns |
| 773 | 1 | 8 | |g volume:293 |g year:2018 |g number:7 |g day:16 |g month:02 |g pages:2597-2605 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1074/jbc.M117.813667 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 293 |j 2018 |e 7 |b 16 |c 02 |h 2597-2605 | ||