Limited proteolysis in the investigation of beta2-microglobulin amyloidogenic and fibrillar states.

Amyloid fibrils of patients treated with regular haemodialysis essentially consists of β2-microglobulin (β2-m) and its truncated species ΔN6β2-m lacking six residues at the amino terminus. The truncated fragment shows a higher propensity to self-aggregate and constitutes an excellent candidate for the analysis of a protein in the amyloidogenic conformation. The surface topology and the conformational analysis of native β2-m and the truncated ΔN6β2-m species both in the soluble and in the fibrillar forms were investigated by the limited proteolysis/mass spectrometry strategy. The conformation in solution of a further truncated mutant ΔN3β2-m lacking three residues at the N-terminus was also examined. This approach appeared particularly suited to investigate the regions that are solvent-exposed, or flexible enough to be accessible to protein-protein interactions and to describe the conformation of transient intermediates. Moreover, proteolysis experiments can also be tailored to investigate amyloid fibrils by discriminating the protein regions constituting the unaccessible core of the fibrils and those still flexible and exposed to the solvent. Although native β2-m and ΔN3β2-m shared essentially the same conformation, significative structural differences exist between the native and the ΔN6β2-m proteins in solution with major differences located at the end moiety of strand V and subsequent loop with strand VI and at both the N- and C-termini of the proteins. On the contrary, an identical distribution of preferential proteolytic sites was observed in both proteins in the fibrillar state, which was nearly superimposible to that observed for the soluble form of ΔN6β2-m. These data revealed that synthetic fibrils essentially consists of an unaccessible core comprising residues 20-87 of the β2-m protein with exposed and flexible N- and C-terminal ends. Moreover, proteolytic cleavages observed in vitro at Lys 6 and Lys 19 reproduce specific cleavages that have to take place in vivo to generate the truncated forms of β2-m occurring in natural fibrils. On the basis of these results, a molecular mechanism for fibril formation has been propose

Universality of three-body systems in 2D: parametrization of the bound states energies

Universal properties of mass-imbalanced three-body systems in 2D are studied using zero-range interactions in momentum space. The dependence of the three-particle binding energy on the parameters (masses and two-body energies) is highly non-trivial even in the simplest case of two identical particles and a distinct one. This dependence is parametrized for ground and excited states in terms of {\itshape supercircles} functions in the most general case of three distinguishable particles.Comment: 3 pages, 1 figure, published versio

Primary cosmic ray spectrum in the 10 to the 12th power - 10 to the 16th power eV energy range from the NUSEX experiment

A primary cosmic ray spectrum was derived which fits both experimental multiple muon rates and the all-nucleon flux derived from the single muon intensities underground. In the frame of the interaction model developed by Gaisser, Elbert and Stanev, it is possible to reproduce NUSEX muon data with a primary composition in which the iron spectrum is only slightly flatter than the proton one. This result rules out the popular idea that the primary composition varies drastically with increasing energy, leading to the dominance of heavier nuclei at energies 10 to the 15th power to 10 to the 16th power eV