Department of Applied Chemistry and Recombinant Protein Expression Center, Sejong University, Seoul 143-747, Korea Received September 23, 2002Reactive oxygen species (ROS) have been investigated to have pivotal roles on amyloidogenecity of β-amyloidpeptide (Aβ), the major component of senile plaques in Alzheimer's disease (AD) brain. Addition of radicalscavengers is one of the on-going strategies for therapeutic treatment for AD patients. Hsp104 protein includingtwo ATP binding sites from Saccharomyces cerevisiae , as a molecular chaperone, was known to function as aprotector of ROS generation when exposed to oxidative stress in our previous study. This observation has ledus to investigate Hsp104 protein as a molecular mediator of A β aggregation in this study. We have developeda new way of expression for Hsp104 protein using GST-fusion tag. As we expected, formation of A β aggregatewas protected by wild type Hsp104 protein, but not by the two ATP-binding site mutant, based on Thioflavin-T fluorescence. Interestingly, Hsp104 protein was observed to keep A β from forming aggregates independentof ATP binding. On the other hand, disaggregation of Aβ aggregates by wild type Hsp104 was totallydependent on the presence of ATP. On the other hand, mutant Hsp104 with two ATP binding sites alteredexhibited no inhibition. Another effective antioxidant, hydrazine analogs of curcumin were also effective in A βfibrilization as protectors against oxidative stress. Based on these observations we conclude that Hsp104 andcurcumin derivatives, as protectors of oxidative stress, inhibit Aβ aggregation in vitro and can be candidatesfor therapeutic approaches in cure of some neurodegenerative diseases.Key Words : Hsp104, Antioxidants, Curcumin, Aβ aggregationIntroductionSeveral neurodegenerative diseases, such as Alzheimer’sDisease (AD), Parkinson Disease (PD) and Huntingtondisease (HD), result from accumulation of damaged proteinsinduced by oxidative stress.