Unfolding, aggregation, and seeded amyloid formation of lysine-58-cleaved beta 2-microglobulin.

Niels H H Heegaard; Thomas J D Jørgensen; Noémi Rozlosnik; Dorthe B Corlin; Jesper S Pedersen; Anna G Tempesta; Peter Roepstorff; Rogert Bauer; Mogens H Nissen

doi:10.1021/bi047594t

Unfolding, aggregation, and seeded amyloid formation of lysine-58-cleaved beta 2-microglobulin.

Niels H H Heegaard, Thomas J D Jørgensen, Noémi Rozlosnik, Dorthe B Corlin, Jesper S Pedersen, Anna G Tempesta, Peter Roepstorff, Rogert Bauer, Mogens H Nissen

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

58 Citationer (Scopus)

Abstract

Udgivelsesdato: 2005-Mar-22

Originalsprog	Engelsk
Tidsskrift	Biochemistry
Vol/bind	44
Udgave nummer	11
Sider (fra-til)	4397-407
Antal sider	10
ISSN	0006-2960
DOI	https://doi.org/10.1021/bi047594t
Status	Udgivet - 2005

Bibliografisk note

Keywords: Amyloid; Binding Sites; Chromatography, Gel; Congo Red; Deuterium Exchange Measurement; Electrophoresis, Capillary; Fluorescent Dyes; Humans; Hydrolysis; Lysine; Microscopy, Atomic Force; Protein Conformation; Protein Folding; Spectrometry, Fluorescence; Spectrometry, Mass, Electrospray Ionization; Temperature; Thiazoles; Time Factors; beta 2-Microglobulin

Adgang til dokumentet

10.1021/bi047594t

Citationsformater

@article{10115a30b93911ddae57000ea68e967b,

title = "Unfolding, aggregation, and seeded amyloid formation of lysine-58-cleaved beta 2-microglobulin.",

abstract = "Beta(2)-microglobulin (beta(2)m) is the amyloidogenic protein in dialysis-related amyloidosis, but the mechanisms underlying beta(2)m fibrillogenesis in vivo are largely unknown. We study a structural variant of beta(2)m that has been linked to cancer and inflammation and may be present in the circulation of dialysis patients. This beta(2)m variant, DeltaK58-beta(2)m, is a disulfide-linked two-chain molecule consisting of amino acid residues 1-57 and 59-99 of intact beta(2)m, and we here demonstrate and characterize its decreased conformational stability as compared to wild-type (wt) beta(2)m. Using amide hydrogen/deuterium exchange monitored by mass spectrometry, we show that DeltaK58-beta(2)m has increased unfolding rates compared to wt-beta(2)m and that unfolding is highly temperature dependent. The unfolding rate is 1 order of magnitude faster in DeltaK58-beta(2)m than in wt-beta(2)m, and at 37 degrees C the half-time for unfolding is more than 170-fold faster than at 15 degrees C. Conformational changes are also reflected by a very prominent Congo red binding of DeltaK58-beta(2)m at 37 degrees C, by the evolution of thioflavin T fluorescence, and by changes in intrinsic fluorescence. After a few days at 37 degrees C, in contrast to wt-beta(2)m, DeltaK58-beta(2)m forms well-defined high molecular weight aggregates that are detected by size-exclusion chromatography. Atomic force microscopy after seeding with amyloid-beta(2)m fibrils under conditions that induce minimal fibrillation in wt-beta(2)m shows extensive amyloid fibrillation in DeltaK58-beta(2)m samples. The results highlight the instability and amyloidogenicity under near physiological conditions of a slightly modified beta(2)m variant generated by limited proteolysis and illustrate stages of amyloid formation from early conformational variants to overt fibrillation.",

author = "Heegaard, {Niels H H} and J{\o}rgensen, {Thomas J D} and No{\'e}mi Rozlosnik and Corlin, {Dorthe B} and Pedersen, {Jesper S} and Tempesta, {Anna G} and Peter Roepstorff and Rogert Bauer and Nissen, {Mogens H}",

note = "Keywords: Amyloid; Binding Sites; Chromatography, Gel; Congo Red; Deuterium Exchange Measurement; Electrophoresis, Capillary; Fluorescent Dyes; Humans; Hydrolysis; Lysine; Microscopy, Atomic Force; Protein Conformation; Protein Folding; Spectrometry, Fluorescence; Spectrometry, Mass, Electrospray Ionization; Temperature; Thiazoles; Time Factors; beta 2-Microglobulin",

year = "2005",

doi = "10.1021/bi047594t",

language = "English",

volume = "44",

pages = "4397--407",

journal = "Biochemistry",

issn = "0006-2960",

publisher = "American Chemical Society",

number = "11",

}

TY - JOUR

T1 - Unfolding, aggregation, and seeded amyloid formation of lysine-58-cleaved beta 2-microglobulin.

AU - Heegaard, Niels H H

AU - Jørgensen, Thomas J D

AU - Rozlosnik, Noémi

AU - Corlin, Dorthe B

AU - Pedersen, Jesper S

AU - Tempesta, Anna G

AU - Roepstorff, Peter

AU - Bauer, Rogert

AU - Nissen, Mogens H

N1 - Keywords: Amyloid; Binding Sites; Chromatography, Gel; Congo Red; Deuterium Exchange Measurement; Electrophoresis, Capillary; Fluorescent Dyes; Humans; Hydrolysis; Lysine; Microscopy, Atomic Force; Protein Conformation; Protein Folding; Spectrometry, Fluorescence; Spectrometry, Mass, Electrospray Ionization; Temperature; Thiazoles; Time Factors; beta 2-Microglobulin

PY - 2005

Y1 - 2005

N2 - Beta(2)-microglobulin (beta(2)m) is the amyloidogenic protein in dialysis-related amyloidosis, but the mechanisms underlying beta(2)m fibrillogenesis in vivo are largely unknown. We study a structural variant of beta(2)m that has been linked to cancer and inflammation and may be present in the circulation of dialysis patients. This beta(2)m variant, DeltaK58-beta(2)m, is a disulfide-linked two-chain molecule consisting of amino acid residues 1-57 and 59-99 of intact beta(2)m, and we here demonstrate and characterize its decreased conformational stability as compared to wild-type (wt) beta(2)m. Using amide hydrogen/deuterium exchange monitored by mass spectrometry, we show that DeltaK58-beta(2)m has increased unfolding rates compared to wt-beta(2)m and that unfolding is highly temperature dependent. The unfolding rate is 1 order of magnitude faster in DeltaK58-beta(2)m than in wt-beta(2)m, and at 37 degrees C the half-time for unfolding is more than 170-fold faster than at 15 degrees C. Conformational changes are also reflected by a very prominent Congo red binding of DeltaK58-beta(2)m at 37 degrees C, by the evolution of thioflavin T fluorescence, and by changes in intrinsic fluorescence. After a few days at 37 degrees C, in contrast to wt-beta(2)m, DeltaK58-beta(2)m forms well-defined high molecular weight aggregates that are detected by size-exclusion chromatography. Atomic force microscopy after seeding with amyloid-beta(2)m fibrils under conditions that induce minimal fibrillation in wt-beta(2)m shows extensive amyloid fibrillation in DeltaK58-beta(2)m samples. The results highlight the instability and amyloidogenicity under near physiological conditions of a slightly modified beta(2)m variant generated by limited proteolysis and illustrate stages of amyloid formation from early conformational variants to overt fibrillation.

AB - Beta(2)-microglobulin (beta(2)m) is the amyloidogenic protein in dialysis-related amyloidosis, but the mechanisms underlying beta(2)m fibrillogenesis in vivo are largely unknown. We study a structural variant of beta(2)m that has been linked to cancer and inflammation and may be present in the circulation of dialysis patients. This beta(2)m variant, DeltaK58-beta(2)m, is a disulfide-linked two-chain molecule consisting of amino acid residues 1-57 and 59-99 of intact beta(2)m, and we here demonstrate and characterize its decreased conformational stability as compared to wild-type (wt) beta(2)m. Using amide hydrogen/deuterium exchange monitored by mass spectrometry, we show that DeltaK58-beta(2)m has increased unfolding rates compared to wt-beta(2)m and that unfolding is highly temperature dependent. The unfolding rate is 1 order of magnitude faster in DeltaK58-beta(2)m than in wt-beta(2)m, and at 37 degrees C the half-time for unfolding is more than 170-fold faster than at 15 degrees C. Conformational changes are also reflected by a very prominent Congo red binding of DeltaK58-beta(2)m at 37 degrees C, by the evolution of thioflavin T fluorescence, and by changes in intrinsic fluorescence. After a few days at 37 degrees C, in contrast to wt-beta(2)m, DeltaK58-beta(2)m forms well-defined high molecular weight aggregates that are detected by size-exclusion chromatography. Atomic force microscopy after seeding with amyloid-beta(2)m fibrils under conditions that induce minimal fibrillation in wt-beta(2)m shows extensive amyloid fibrillation in DeltaK58-beta(2)m samples. The results highlight the instability and amyloidogenicity under near physiological conditions of a slightly modified beta(2)m variant generated by limited proteolysis and illustrate stages of amyloid formation from early conformational variants to overt fibrillation.

U2 - 10.1021/bi047594t

DO - 10.1021/bi047594t

M3 - Journal article

C2 - 15766269

SN - 0006-2960

VL - 44

SP - 4397

EP - 4407

JO - Biochemistry

JF - Biochemistry

IS - 11

ER -