Investigating the influences of redox buffer compositions on the amyloid fibrillogenesis of hen egg-white lysozyme.
More than twenty different human proteins have been found to fold abnormally resulting in the formation of pathological deposits and several lethal degenerative diseases. Despite extensive investigations on amyloid fibril formation, the detailed molecular mechanism remained rather elusive. The present study is aimed at exploring the effect of the ratio of cysteine and cystine in the buffer on the fibrillation of hen egg-white lysozyme. Our results revealed that the inhibition of lysozyme amyloid formation by cysteine in the redox buffer followed a concentration-dependent fashion. Cystine, the oxidized form of cysteine, nevertheless, did not influence the final level of fibrillation although it lengthened the lag period of fibril formation. Moreover, the effect of the ratio of cysteine to cystine in the buffer on the fibrillogenesis of hen lysozyme was found to be greatly associated with the formation of mixed disulfide derivatives. Finally, a possible reaction mechanism was proposed to explain our experimental results. Our study shows that the concentration of mixed disulfide derivative was inversely correlated with the level of lysozyme fibrillogenesis. The results from this work may aid in comprehending the molecular mechanism(s) of amyloid fibrillogenesis for disulfide bonded proteins and the development of effective therapeutics for amyloidogenic diseases.
Wang
Steven S-S
SS
Department of Chemical Engineering, National Taiwan University, No.1, Sec.4, Roosevelt Rd., Taipei 10617, Taiwan. sswang@ntu.edu.tw
Liu
Kuan-Nan
KN
Wu
Chia-Hung
CH
Lai
Jun-Kun
JK
D000682:Amyloid / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D002021:Buffers; D003545:Cysteine / Q000737:chemistry; D003553:Cystine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D007700:Kinetics; D046529:Microscopy, Electron, Transmission; D009113:Muramidase / Q000187:drug effects / Q000378:metabolism*; D010084:Oxidation-Reduction
D000682:Amyloid; D002021:Buffers; D004220:Disulfides; D003553:Cystine; C421864:hen egg lysozyme; D009113:Muramidase; D003545:Cysteine