A mass spectrometric approach to study the interaction of amyloid β peptides with human α-2-macroglobulin

We used MALDI-MS to study the interaction of amyloid β (Aβ) peptides with alpha-2-macroglobulin (α2M). The binding of amyloid beta (Aβ) peptides to alpha-2-macroglobulin (α2M) was found to inhibit the ability of trypsin to cleave out the peptide α2M 705-715 (Pep-α2M) from α2M. This was observed with both purified α2M and α2M in human serum. We found that Aβ 1-38, Aβ1-40, and Aβ 1-42, all inhibit the interaction of α2M with trypsin, with inhibition rate independent of the length of the Aβ peptide. Further, we show that for complete inhibition, two peptide molecules must be attached to one α2M molecule; one for each of its two subunits. A region was revealed within the Aβ sequence, in which proteolytic cleavage (Lys-28) and oxidation (Met-35) lead to a loss of their ability to inhibit the interaction of trypsin with α2M. Furthermore, we show that after the formation of a trypsin complex with α2M and cleavage of α2M to produce the α2M 705-715, Aβ peptides continue to bind to the protein in the same proportions. However, Aβ peptides treated with DMSO lost their ability to bind to α2M and thereby to inhibit the interaction of trypsin with α2M. While maintaining their primary structure, such an effect can be explained only by conformational changes in the peptides, suggesting the possibility to use our analytical approach to distinguish between conformational isomers of Aβ peptides.