Bela2-microglobuline amyloidosis: Fibrillogenesis of natural and recombinant human beta2-microglobulines


Β2-Microglobulin ( β2М) is noncovalently associated with alpha chain of the class I major histocompatibility complex. The protein is removed from blood by degradation in the proximal tubule of the kidney. β2M cannot be removed from the blood by the kidney or the dialysis membrane in patients undergoing haemodialysis. As a consequence of increased β2M concentrations, partial unfolding of β2M is believed to be prerequisite to its assembly into β2M amyloid fibrils. β2M amyloidosis is a common complication associated with long-term hemodialysis. Mechanism of fibril formation in physiological conditions is poorly understood. Simulation in vitro is required for elucidation of its mechanism and for searching after factors that can induce or inhibit fibrillogenesis. For these purpose we have devised the method to obtain β2M from dialysate of patients undergoing long-term hemodialysis. We have constructed expression plasmid for production of recombinant human β2M in E.coli with subsequent fast and easy purification. Recombinant fusion protein of β2M with green fluorescent protein was also produced. These recombinant proteins do not form inclusion bodies and can be extracted directly from the cell lysate. Electron microscopy and atomic force microscopy have shown presence of fibrils formed in vitro from the three β2M variants. Using thioflavin-T binding we have determined that β2M purified from dialysate and recombinant β2M can form amyloid fibrils.

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