Amyloid protofilaments from the calcium-binding protein equine lysozyme: Formation of ring and linear structures depends on pH and metal ion concentration

被引:94
作者
Malisauskas, M
Zamotin, V
Jass, J
Noppe, W
Dobson, CM
Morozova-Roche, LA [1 ]
机构
[1] Umea Univ, Dept Biochem, SE-90187 Umea, Sweden
[2] Umea Univ, Dept Microbiol, SE-90187 Umea, Sweden
[3] Katholieke Univ Leuven, Interdisciplinary Res Ctr, B-8500 Kortrijk, Belgium
[4] Univ Cambridge, Dept Chem, Cambridge CB2 1EW, England
基金
英国惠康基金;
关键词
amyloidosis; lysozyme; protofilament; circularisation; acidic hydrolysis;
D O I
10.1016/S0022-2836(03)00551-5
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
The calcium-binding equine lysozyme has been found to undergo conversion into amyloid fibrils during incubation in solution at acidic pH. At pH 4.5 and 57 degreesC, where equine lysozyme forms a partially unfolded molten globule state, the protein forms protofilaments with a width of ca. 2 nm. In the absence of Ca2+ the protofilaments are present as annular structures with a diameter of 40-50 nm. In the presence of 10 mM CaCl2 the protofilaments of equine lysozyme are straight or curved; they can assemble into thicker threads, but they do not appear to undergo circularisation. At pH 2.0, where the protein is more destabilised compared to pH 4.5, fibril formation occurs at 37 degreesC and 57 degreesC. At pH 2.0, both ring-shaped and linear protofilaments are formed, in which periodic repeats of ca 35 nm can be distinguished clearly. The rings constitute about 10% of all fibrillar species under these conditions and they are characterised by a larger diameter of 70-80 nm. All the structures bind Congo red and thioflavine T in a manner similar to fibrils associated with a variety of amyloid diseases. At pH 2.0, fibril formation is accompanied by some acidic hydrolysis, producing specific fragmentation of the protein, leading to the accumulation of two peptides in particular, consisting of residues 1-80 and 54-125. At the initial stages of incubation, however, full-length equine lysozyme represents the dominant species within the fibrils. We propose that the ring-shaped structures observed here, and in the case of disease-associated proteins such as alpha-synuclein, could be a second generic type of amyloid structure in addition to the more common linear fibrils. (C) 2003 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:879 / 890
页数:12
相关论文
共 71 条
[1]  
Blanch EW, 2000, BIOPOLYMERS, V57, P235, DOI 10.1002/1097-0282(2000)57:4&lt
[2]  
235::AID-BIP5&gt
[3]  
3.0.CO
[4]  
2-H
[5]   Is polyproline II helix the killer conformation? A Raman optical activity study of the amyloidogenic prefibrillar intermediate of human lysozyme [J].
Blanch, EW ;
Morozova-Roche, LA ;
Cochran, DAE ;
Doig, AJ ;
Hecht, L ;
Barron, LD .
JOURNAL OF MOLECULAR BIOLOGY, 2000, 301 (02) :553-563
[6]   Instability, unfolding and aggregation of human lysozyme variants underlying amyloid fibrillogenesis [J].
Booth, DR ;
Sunde, M ;
Bellotti, V ;
Robinson, CV ;
Hutchinson, WL ;
Fraser, PE ;
Hawkins, PN ;
Dobson, CM ;
Radford, SE ;
Blake, CCF ;
Pepys, MB .
NATURE, 1997, 385 (6619) :787-793
[7]   Inherent toxicity of aggregates implies a common mechanism for protein misfolding diseases [J].
Bucciantini, M ;
Giannoni, E ;
Chiti, F ;
Baroni, F ;
Formigli, L ;
Zurdo, JS ;
Taddei, N ;
Ramponi, G ;
Dobson, CM ;
Stefani, M .
NATURE, 2002, 416 (6880) :507-511
[8]   Mechanistic studies of the folding of human lysozyme and the origin of amyloidogenic behavior in its disease-related variants [J].
Canet, D ;
Sunde, M ;
Last, AM ;
Miranker, A ;
Spencer, A ;
Robinson, CV ;
Dobson, CM .
BIOCHEMISTRY, 1999, 38 (20) :6419-6427
[9]   Ultrastructural organization of amyloid fibrils by atomic force microscopy [J].
Chamberlain, AK ;
MacPhee, CE ;
Zurdo, J ;
Morozova-Roche, LA ;
Hill, HAO ;
Dobson, CM ;
Davis, JJ .
BIOPHYSICAL JOURNAL, 2000, 79 (06) :3282-3293
[10]   Furin initiates gelsolin familial amyloidosis in the Golgi through a defect in Ca2+ stabilization [J].
Chen, CD ;
Huff, ME ;
Matteson, J ;
Page, L ;
Phillips, R ;
Kelly, JW ;
Balch, WE .
EMBO JOURNAL, 2001, 20 (22) :6277-6287