%I mi
%T Analysis of the Native Structure, Stability and Aggregation of Biotinylated Human Lysozyme
%X Fibril formation by mutational variants of human lysozyme is associated with a fatal form of hereditary non-neuropathic systemic amyloidosis. Defining the mechanistic details of lysozyme aggregation is of crucial importance for understanding the origin and progression of this disease and related misfolding conditions. In this study, we show that a biotin moiety can be introduced site-specifically at Lys33 of human lysozyme. We demonstrate, using biophysical techniques, that the structure and stability of the native-state of the protein are not detectably altered by this modification, and that the ability to form amyloid fibrils is unchanged. By taking advantage of biotin-avidin interactions, we show that super-resolution fluorescence microscopy can generate detailed images of the mature fibrils. This methodology can readily enable the introduction of additional probes into the protein, thereby providing the means through which to understand, in detail, the nature of the aggregation process of lysozyme and its variants under a variety of conditions. Š 2012 Ahn et al.
%D 2012
%L publicatio23188
%V 7
%P Terjedelem: 10 p-AzonosĂ­tĂł: e50192
%N 11
%J PLOS ONE
%O Department of Chemistry, University of Cambridge, Cambridge, United Kingdom            
            Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge, United Kingdom            
            National Physics Laboratory, Teddington, United Kingdom            
            Cited By :19            
            Export Date: 25 May 2020            
            Correspondence Address: Kumita, J. R.; Department of Chemistry, University of Cambridge, Cambridge, United Kingdom; email: jrk38@cam.ac.uk            
            Chemicals/CAS: amyloid, 11061-24-8; biotin, 58-85-5; lysine, 56-87-1, 6899-06-5, 70-54-2; lysozyme, 9001-63-2; Amyloid; Avidin, 1405-69-2; Muramidase, 3.2.1.17; lysozyme C, human, 3.2.1.17
%A  Ahn Minkoo
%A  Genst, de Edwin
%A  Kaminski Schierle Gabriele S.
%A  ErdĂŠlyi MiklĂłs
%A  Kaminski Clemens F.
%A  Dobson Melanie J.
%A  Kumita Janet R.
%R MTMT:2206013 10.1371/journal.pone.0050192