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Lysine 58-cleaved beta 2-microglobulin is not detectable by 2D electrophoresis in ex vivo amyloid fibrils of two patients affected by dialysis-related amyloidosis

Giorgetti, S; Stoppini, M; Tennent, GA; Relini, A; Marchese, L; Raimondi, S; Monti, M; ... Bellotti, V; + view all (2007) Lysine 58-cleaved beta 2-microglobulin is not detectable by 2D electrophoresis in ex vivo amyloid fibrils of two patients affected by dialysis-related amyloidosis. PROTEIN SCI , 16 (2) 343 - 349. 10.1110/ps.062563507.

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Abstract

The lysine 58 cleaved and truncated variant of beta(2)-microglobulin (Delta K58-beta 2m) is conformationally unstable and present in the circulation of a large percentage of patients on chronic hemodialysis, suggesting that it could play a role in the beta 2-microglobulin (beta 2m) amyloid fibrillogenesis associated with dialysis-related amyloidosis (DRA). However, it has yet to be detected in the amyloid deposits of such patients. Here, we extracted amyloid fibrils, without denaturation or additional purification, from different amyloidotic tissues of two unrelated individuals suffering from DRA, and characterized them by high-sensitivity bidimensional gel electrophoresis (2D-PAGE), immunoblotting, MALDI time-of-flight mass spectrometry, and protein sequencing. To confirm whether or not this species could be identified by our proteomic approaches, we mapped its location in 2D-PAGE, in mixtures of pure Delta K58-beta 2m, and extracts of amyloid fibrils from patients, to a discrete region of the gel distinct from other isoforms of beta 2m. Using this approach, the two known principal isoforms found in beta 2m amyloid were identified, namely, the full-length protein and the truncated species lacking six N-terminal amino acid residues (Delta N6-beta 2m). In contrast, we found no evidence for the presence of Delta K58-beta 2m.

Type: Article
Title: Lysine 58-cleaved beta 2-microglobulin is not detectable by 2D electrophoresis in ex vivo amyloid fibrils of two patients affected by dialysis-related amyloidosis
DOI: 10.1110/ps.062563507
Keywords: dialysis related amyloidosis, amyloid fibrils, proteolized variants of beta 2-microglobulin, proteomics, CHRONIC-HEMODIALYSIS, PROTEIN AGGREGATION, BETA(2)-MICROGLOBULIN, SERUM, FRAGMENTS, VARIANTS
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine > Inflammation
URI: http://discovery.ucl.ac.uk/id/eprint/90541
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