A standardized and biocompatible preparation of aggregate-free amyloid beta peptide for biophysical and biological studies of Alzheimers disease

被引：102

作者：

Broersen, Kerensa ^{[1
,2
]}

Jonckheere, Wim ^{[1
,2
]}

Rozenski, Jef ^{[3
]}

Vandersteen, Annelies ^{[1
,2
]}

Pauwels, Kris ^{[4
]}

Pastore, Annalisa ^{[4
]}

Rousseau, Frederic ^{[1
,2
]}

Schymkowitz, Joost ^{[1
,2
]}

机构：

[1] VIB, SWITCH Lab, B-1050 Brussels, Belgium

[2] VUB, B-1050 Brussels, Belgium

[3] Katholieke Univ Leuven, Rega Inst Med Res, Med Chem Lab, B-3000 Louvain, Belgium

[4] Natl Inst Med Res, MRC, London NW7 1AA, England

来源：

PROTEIN ENGINEERING DESIGN & SELECTION | 2011年 / 24卷 / 09期

关键词：

aggregation; amyloid beta peptide; biocompatibility; in vitro studies; solubilization; GAMMA-SECRETASE ACTIVITY; PRECURSOR PROTEIN; FIBRILS; HEXAFLUOROISOPROPANOL; NEUROTOXICITY; PRESENILIN-1; A-BETA(1-42); MUTATIONS; SPECTRA; FIBRILLOGENESIS;

D O I：

10.1093/protein/gzr020

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We provide a validated and rapid protocol for the solubilization of amyloid -peptide (A). This procedure involves sequential solubilization using structure-breaking organic solvents hexafluoroisopropanol and DMSO followed by column purification. The low solubility and tendency of A to aggregate considerably impede the in vitro handling and biophysical or biological investigation of A, despite the interest in this peptide because of its implication in Alzheimers disease. The main advantage of the proposed protocol over others is that it results in standardized aggregate-free A peptide samples that are biocompatible for cell culture studies and yield reproducible aggregation kinetics and cytotoxicities. This three-step protocol also enables the co-solubilization of the longer A42 variant with A40 in ratios relevant to Alzheimers disease.

引用

页码：743 / 750

页数：8

共 48 条

[1] THE PROGRAM XEASY FOR COMPUTER-SUPPORTED NMR SPECTRAL-ANALYSIS OF BIOLOGICAL MACROMOLECULES [J].

BARTELS, C ;

XIA, TH ;

BILLETER, M ;

GUNTERT, P ;

WUTHRICH, K .

JOURNAL OF BIOMOLECULAR NMR, 1995, 6 (01) :1-10

[2] Presenilin clinical mutations can affect γ-secretase activity by different mechanisms [J].

Bentahir, M ;

Nyabi, O ;

Verhamme, J ;

Tolia, A ;

Horré, K ;

Wiltfang, J ;

Esselmann, H ;

De Strooper, B .

JOURNAL OF NEUROCHEMISTRY, 2006, 96 (03) :732-742

[3] Amyloid β-protein:: Monomer structure and early aggregation states of Aβ42 and its Pro19 alloform [J].

Bernstein, SL ;

Wyttenbach, T ;

Baumketner, A ;

Shea, JE ;

Bitan, G ;

Teplow, DB ;

Bowers, MT .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2005, 127 (07) :2075-2084

[4]

Bitan Gal, 2004, V299, P3

[5]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[6] Trifluoroethanol and colleagues: cosolvents come of age. Recent studies with peptides and proteins [J].

Buck, M .

QUARTERLY REVIEWS OF BIOPHYSICS, 1998, 31 (03) :297-355

[7] Alzheimer's amyloid β-peptide (1-42):: involvement of methionine residue 35 in the oxidative stress and neurotoxicity properties of this peptide [J].

Butterfield, DA ;

Bush, AI .

NEUROBIOLOGY OF AGING, 2004, 25 (05) :563-568

[8] Antiparallel β-sheet: a signature structure of the oligomeric amyloid β-peptide [J].

Cerf, Emilie ;

Sarroukh, Rabia ;

Tamamizu-Kato, Shiori ;

Breydo, Leonid ;

Derclaye, Sylvie ;

Dufrene, Yves F. ;

Narayanaswami, Vasanthy ;

Goormaghtigh, Erik ;

Ruysschaert, Jean-Marie ;

Raussens, Vincent .

BIOCHEMICAL JOURNAL, 2009, 421 :415-423

[9] Rationalization of the effects of mutations on peptide and protein aggregation rates [J].

Chiti, F ;

Stefani, M ;

Taddei, N ;

Ramponi, G ;

Dobson, CM .

NATURE, 2003, 424 (6950) :805-808

[10] Self-assembly of Aβ1-42 into globular neurotoxins [J].

Chromy, BA ;

Nowak, RJ ;

Lambert, MP ;

Viola, KL ;

Chang, L ;

Velasco, PT ;

Jones, BW ;

Fernandez, SJ ;

Lacor, PN ;

Horowitz, P ;

Finch, CE ;

Krafft, GA ;

Klein, WL .

BIOCHEMISTRY, 2003, 42 (44) :12749-12760

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