Human Liver Oxidative Stress And Oxidation Of A Preliminary Study Of Transcriptional Networks

Since liver is the major metabolism center for human being, ROS will be produced as a natural by-product of it's actively metabolism processes. The excessive generation of ROS and/or a defect or deficiency in the anti-oxidant defense system can result in a state called oxidative stress. Alcohol abuse, increased drug intake or the metabolism of excessive free fatty acids in obese patients also cause oxidative stress in the hepatocytes, which has been shown to contribute to various liver diseases. Since oxidative stress is easily induced in hepatocyte, the cellular oxidative stress response and redox regulation mechanisms must play critical roles in the protection of liver from oxidative damage. But their protection roles and mechanisms in human liver have not been systematically researched.The elucidation of protein-protein interaction patterns in large-scale provides an important basic data set in the functional analysis of the proteome. In this dissertation, we undertake a primary research on the oxidative stress response and redox regulation network in human liver by large-scale Y2H library screening with proteins involved in such process.Twenty seven baits were successfully screened against human liver cDNA library. Over 528 candidate colonies were isolated initially. Their AD-Y identities were determined with interaction sequence tags (ISTs) obtained by sequencing the corresponding polymerase chain reaction (PCR) products. The AD-Y reading frame was verified for each IST. After removal of sequences mapping to the non-coding DNA region and the coding sequence out of AD frame, 223 cDNA fragments were retained in our data set, forming 97 different protein interaction pairs involved 20 baits. Among which 11 interactions are known and 87% of the newly identified interations are unkown. These results may seguesst that there exist many unrecognized members in the oxidative stress response and redox regulation network.To estimate our technical false positive rate, 61 randomly selected interactions were verified by reassessment of the interactions in yeast cells. The total recovery for all the 61 interactions was 52.5%. With these results, it should be reasoned that there exist some technical false positives in our data set. But only 53.3% of the recovery for the known interactions seguests that prominent false negative rate was lied in the retest...