Spontaneous aggregation of folded and soluble native proteins in vivo is still a poorly understood process. A prototypic example is the D76N mutant of beta-2 microglobulin (β2m) that displays an aggressive aggregation propensity. Here we investigate the dynamics of β2m by X-ray crystallography, solid-state NMR, and molecular dynamics simulations to unveil the effects of the D76N mutation. Taken together, our data highlight the presence of minor disordered substates in crystalline β2m. The destabilization of the outer strands of D76N β2m accounts for the increased aggregation propensity. Furthermore, the computational modeling reveals a network of interactions with residue D76 as a keystone: this model allows predicting the stability of several point mutants. Overall, our study shows how the study of intrinsic dynamics in crystallo can provide crucial answers on protein stability and aggregation propensity. The comprehensive approach here presented may well be suited for the study of other folded amyloidogenic proteins.

Conformational dynamics in crystals reveal the molecular bases for D76N beta-2 microglobulin aggregation propensity / T. Le Marchand, M. de Rosa, N. Salvi, B.M. Sala, L.B. Andreas, E. Barbet-Massin, P. Sormanni, A. Barbiroli, R. Porcari, C. Sousa Mota, D. de Sanctis, M. Bolognesi, L. Emsley, V. Bellotti, M. Blackledge, C. Camilloni, G. Pintacuda, S. Ricagno. - In: NATURE COMMUNICATIONS. - ISSN 2041-1723. - 9:1(2018), pp. 1658.1-1658.11.

Conformational dynamics in crystals reveal the molecular bases for D76N beta-2 microglobulin aggregation propensity

M. de Rosa
Secondo
;
B.M. Sala;A. Barbiroli;M. Bolognesi;C. Camilloni
;
S. Ricagno
Ultimo
2018

Abstract

Spontaneous aggregation of folded and soluble native proteins in vivo is still a poorly understood process. A prototypic example is the D76N mutant of beta-2 microglobulin (β2m) that displays an aggressive aggregation propensity. Here we investigate the dynamics of β2m by X-ray crystallography, solid-state NMR, and molecular dynamics simulations to unveil the effects of the D76N mutation. Taken together, our data highlight the presence of minor disordered substates in crystalline β2m. The destabilization of the outer strands of D76N β2m accounts for the increased aggregation propensity. Furthermore, the computational modeling reveals a network of interactions with residue D76 as a keystone: this model allows predicting the stability of several point mutants. Overall, our study shows how the study of intrinsic dynamics in crystallo can provide crucial answers on protein stability and aggregation propensity. The comprehensive approach here presented may well be suited for the study of other folded amyloidogenic proteins.
NMR chemical-shifts; proton-detected NMR; rational design; amyloid fibrils; protein; state; relaxation; model; assignment; backbone
Settore BIO/10 - Biochimica
Settore FIS/07 - Fisica Applicata(Beni Culturali, Ambientali, Biol.e Medicin)
From basic knowledge on the dynamics of b2-m self aggregation to the discovery of new inhibitors of amyloidogenesis
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/571567
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