Aβ Protofibrils Possess a Stable Core Structure Resistant to Hydrogen Exchange

Indu Kheterpal, Hilal A. Lashuel, Dean M. Hartley, Thomas Walz, Peter T. Lansbury, Ronald Wetzel

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

Protofibrils are transient structures observed during in vitro formation of mature amyloid fibrils and have been implicated as the toxic species responsible for cell dysfunction and neuronal loss in Alzheimer's disease (AD) and other protein aggregation diseases. To better understand the roles of protofibrils in amyloid assembly and Alzheimer's disease, we characterized secondary structural features of these heterogeneous and metastable assembly intermediates. We chromatographically isolated different size populations of protofibrils from amyloid assembly reactions of Aβ(1-40), both wild type and the Arctic variant associated with early onset familial AD, and exposed them to hydrogen-deuterium exchange analysis monitored by mass spectrometry (HX-MS). We show that HX-MS can distinguish among unstructured monomer, protofibrils, and fibrils by their different protection patterns. We find that about 40% of the backbone amide hydrogens of Aβ protofibrils are highly resistant to exchange with deuterium even after 2 days of incubation in aqueous deuterated buffer, implying a very stable, presumably H-bonded, core structure. This is in contrast to mature amyloid fibrils, whose equally stable structure protects about 60% of the backbone amide hydrogens over the same time frame. We also find a surprising degree of specificity in amyloid assembly, in that wild type Aβ is preferentially excluded from both protofibrils and fibrils grown from an equimolar mixture of wild type and Arctic mutant peptides. These and other data are interpreted and discussed in terms of the role of protofibrils in fibril assembly and in disease.

Original languageEnglish
Pages (from-to)14092-14098
Number of pages7
JournalBiochemistry
Volume42
Issue number48
DOIs
Publication statusPublished - 9 Dec 2003
Externally publishedYes

Fingerprint

Amyloid
Hydrogen
Alzheimer Disease
Deuterium
Amides
Poisons
Population Density
Mass spectrometry
Mass Spectrometry
Buffers
Agglomeration
Monomers
Peptides
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kheterpal, I., Lashuel, H. A., Hartley, D. M., Walz, T., Lansbury, P. T., & Wetzel, R. (2003). Aβ Protofibrils Possess a Stable Core Structure Resistant to Hydrogen Exchange. Biochemistry, 42(48), 14092-14098. https://doi.org/10.1021/bi0357816

Aβ Protofibrils Possess a Stable Core Structure Resistant to Hydrogen Exchange. / Kheterpal, Indu; Lashuel, Hilal A.; Hartley, Dean M.; Walz, Thomas; Lansbury, Peter T.; Wetzel, Ronald.

In: Biochemistry, Vol. 42, No. 48, 09.12.2003, p. 14092-14098.

Research output: Contribution to journalArticle

Kheterpal, I, Lashuel, HA, Hartley, DM, Walz, T, Lansbury, PT & Wetzel, R 2003, 'Aβ Protofibrils Possess a Stable Core Structure Resistant to Hydrogen Exchange', Biochemistry, vol. 42, no. 48, pp. 14092-14098. https://doi.org/10.1021/bi0357816
Kheterpal I, Lashuel HA, Hartley DM, Walz T, Lansbury PT, Wetzel R. Aβ Protofibrils Possess a Stable Core Structure Resistant to Hydrogen Exchange. Biochemistry. 2003 Dec 9;42(48):14092-14098. https://doi.org/10.1021/bi0357816
Kheterpal, Indu ; Lashuel, Hilal A. ; Hartley, Dean M. ; Walz, Thomas ; Lansbury, Peter T. ; Wetzel, Ronald. / Aβ Protofibrils Possess a Stable Core Structure Resistant to Hydrogen Exchange. In: Biochemistry. 2003 ; Vol. 42, No. 48. pp. 14092-14098.
@article{9baf5f956872468f82f38bb6a09d60e3,
title = "Aβ Protofibrils Possess a Stable Core Structure Resistant to Hydrogen Exchange",
abstract = "Protofibrils are transient structures observed during in vitro formation of mature amyloid fibrils and have been implicated as the toxic species responsible for cell dysfunction and neuronal loss in Alzheimer's disease (AD) and other protein aggregation diseases. To better understand the roles of protofibrils in amyloid assembly and Alzheimer's disease, we characterized secondary structural features of these heterogeneous and metastable assembly intermediates. We chromatographically isolated different size populations of protofibrils from amyloid assembly reactions of Aβ(1-40), both wild type and the Arctic variant associated with early onset familial AD, and exposed them to hydrogen-deuterium exchange analysis monitored by mass spectrometry (HX-MS). We show that HX-MS can distinguish among unstructured monomer, protofibrils, and fibrils by their different protection patterns. We find that about 40{\%} of the backbone amide hydrogens of Aβ protofibrils are highly resistant to exchange with deuterium even after 2 days of incubation in aqueous deuterated buffer, implying a very stable, presumably H-bonded, core structure. This is in contrast to mature amyloid fibrils, whose equally stable structure protects about 60{\%} of the backbone amide hydrogens over the same time frame. We also find a surprising degree of specificity in amyloid assembly, in that wild type Aβ is preferentially excluded from both protofibrils and fibrils grown from an equimolar mixture of wild type and Arctic mutant peptides. These and other data are interpreted and discussed in terms of the role of protofibrils in fibril assembly and in disease.",
author = "Indu Kheterpal and Lashuel, {Hilal A.} and Hartley, {Dean M.} and Thomas Walz and Lansbury, {Peter T.} and Ronald Wetzel",
year = "2003",
month = "12",
day = "9",
doi = "10.1021/bi0357816",
language = "English",
volume = "42",
pages = "14092--14098",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "48",

}

TY - JOUR

T1 - Aβ Protofibrils Possess a Stable Core Structure Resistant to Hydrogen Exchange

AU - Kheterpal, Indu

AU - Lashuel, Hilal A.

AU - Hartley, Dean M.

AU - Walz, Thomas

AU - Lansbury, Peter T.

AU - Wetzel, Ronald

PY - 2003/12/9

Y1 - 2003/12/9

N2 - Protofibrils are transient structures observed during in vitro formation of mature amyloid fibrils and have been implicated as the toxic species responsible for cell dysfunction and neuronal loss in Alzheimer's disease (AD) and other protein aggregation diseases. To better understand the roles of protofibrils in amyloid assembly and Alzheimer's disease, we characterized secondary structural features of these heterogeneous and metastable assembly intermediates. We chromatographically isolated different size populations of protofibrils from amyloid assembly reactions of Aβ(1-40), both wild type and the Arctic variant associated with early onset familial AD, and exposed them to hydrogen-deuterium exchange analysis monitored by mass spectrometry (HX-MS). We show that HX-MS can distinguish among unstructured monomer, protofibrils, and fibrils by their different protection patterns. We find that about 40% of the backbone amide hydrogens of Aβ protofibrils are highly resistant to exchange with deuterium even after 2 days of incubation in aqueous deuterated buffer, implying a very stable, presumably H-bonded, core structure. This is in contrast to mature amyloid fibrils, whose equally stable structure protects about 60% of the backbone amide hydrogens over the same time frame. We also find a surprising degree of specificity in amyloid assembly, in that wild type Aβ is preferentially excluded from both protofibrils and fibrils grown from an equimolar mixture of wild type and Arctic mutant peptides. These and other data are interpreted and discussed in terms of the role of protofibrils in fibril assembly and in disease.

AB - Protofibrils are transient structures observed during in vitro formation of mature amyloid fibrils and have been implicated as the toxic species responsible for cell dysfunction and neuronal loss in Alzheimer's disease (AD) and other protein aggregation diseases. To better understand the roles of protofibrils in amyloid assembly and Alzheimer's disease, we characterized secondary structural features of these heterogeneous and metastable assembly intermediates. We chromatographically isolated different size populations of protofibrils from amyloid assembly reactions of Aβ(1-40), both wild type and the Arctic variant associated with early onset familial AD, and exposed them to hydrogen-deuterium exchange analysis monitored by mass spectrometry (HX-MS). We show that HX-MS can distinguish among unstructured monomer, protofibrils, and fibrils by their different protection patterns. We find that about 40% of the backbone amide hydrogens of Aβ protofibrils are highly resistant to exchange with deuterium even after 2 days of incubation in aqueous deuterated buffer, implying a very stable, presumably H-bonded, core structure. This is in contrast to mature amyloid fibrils, whose equally stable structure protects about 60% of the backbone amide hydrogens over the same time frame. We also find a surprising degree of specificity in amyloid assembly, in that wild type Aβ is preferentially excluded from both protofibrils and fibrils grown from an equimolar mixture of wild type and Arctic mutant peptides. These and other data are interpreted and discussed in terms of the role of protofibrils in fibril assembly and in disease.

UR - http://www.scopus.com/inward/record.url?scp=0345060507&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0345060507&partnerID=8YFLogxK

U2 - 10.1021/bi0357816

DO - 10.1021/bi0357816

M3 - Article

C2 - 14640676

AN - SCOPUS:0345060507

VL - 42

SP - 14092

EP - 14098

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 48

ER -