A single disulfide bond differentiates aggregation pathways of β2-microglobulin

Deposition of wild-type beta 2-microglobulin (beta 2m) into amyloid fibrils is a complication in patients undergoing long-term hemodialysis. The native beta-sandwich fold of 92m has a highly conserved disulfide bond linking Cys25 and Cys80. Oxidized 92m forms needle-like amyloid fibrils at pH 2.5 in vitro, whereas reduced 92m, at acid pH, in which the intra-chain disulfide bond is disrupted, cannot form typical fibrils. Instead, reduced beta 2m forms thinner and more flexible filaments. To uncover the difference in molecular mechanisms underlying the aggregation of the oxidized and reduced beta 2m, we performed molecular dynamics simulations of beta 2m oligomerization under oxidized and reduced conditions. We show that, consistent with experimental observations, the oxidized beta 2m forms domain-swapped dimer, in which the two proteins exchange their N-terminal segments complementing each other. In contrast, both dimers and trimers, formed by reduced beta 2m, are comprised of parallel B-sheets between monomers and stabilized by the hydrogen bond network along the backbone. The oligomerized monomers are in extended conformations, capable of further aggregation. We find that both reduced and oxidized dimers are thermodynamically less stable than their corresponding monomers, indicating that beta 2m oligomerization is not accompanied by the formation of a thermodynamically stable dimer. Our studies suggest that the different aggregation pathways of oxidized and reduced beta 2m are dictated by the formation of distinct precursor oligomeric species that are modulated by Cys25-Cys8O disulfide-bonds. We propose that the propagation of domain swapping is the aggregation mechanism for the oxidized beta 2m, while "parallel stacking" of partially unfolded beta 2m is the aggregation mechanism for the reduced beta 2m. (c) 2005 Elsevier Ltd. All rights reserved.