Identification of AL proteins from 10 λ-AL amyloidosis patients by mass spectrometry extracted from abdominal fat and heart tissue

Background Systemic AL amyloidosis arises from the misfolding of patient-specific immunoglobulin light chains (LCs). Potential drivers of LC amyloid formation are mutational changes and post-translational modifications (PTMs). However, little information is available on the exact primary structure of the AL proteins and their precursor LCs. Objective We analyse the exact primary structure of AL proteins extracted from 10 lambda AL amyloidosis patients and their corresponding precursor LCs. Materials and Methods By cDNA sequencing of the precursor LC genes in combination with mass spectrometry of the AL proteins, the exact primary structure and PTMs were determined. This information was used to analyse their biochemical properties. Results All AL proteins comprise the V-L and a small part of the C-L with a common C-terminal truncation region. While all AL proteins retain the conserved native disulphide bond of the V-L, we found no evidence for presence of other common PTMs. The analysis of the biochemical properties revealed that the isoelectric point of the V-L is significantly increased due to introduced mutations. Conclusion Our data imply that mutational changes influence the surface charge properties of the V-L and that common proteolytic processes are involved in the generation of the cleavage sites of AL proteins.