Antioxidant Action of Transthyretin in Human Cerebrospinal Fluid.

Yang, Sung Yeul; Chay, Kee Oh; Park, Jong Geun; Ryu, Moon Hee; Hong, Suck Noh; Kim, Soo Han; Ahn, Bong Whan; Lee, Je Hyuk; Lee, Min Wha

Original Article

Journal of Korean Neurosurgical Society 1994;23(4): 375-381.

Sung Yeul Yang, Kee Oh Chay, Jong Geun Park, Moon Hee Ryu, Suck Noh Hong, Soo Han Kim, Bong Whan Ahn, Je Hyuk Lee, Min Wha Lee

¹Department of Biochemistry, Chonnan University Medical School. Kwangju, Korea.
²Department of Neurosurgery, Chonnan University Medical School. Kwangju, Korea.

ABSTRACT

Protective effect of human cerebrospinal fluid antioxidants against enzyme inactivation caused by metal-catalyzed oxidation systems were investigated. When purified glutamine synthetase(GS) was incubated with human cerebrospinal fluid(CSF), the enzyme was progressively inactivated. Catalase and EDTA could inhibit the enzyme inactivation by 50-80%. Small-molecular(Mr<-10,000) fraction of CSF inactivated the exogenous GS, but large-molecular(Mr>-10,000) fraction did not. The GS inactivation by the small-molecular fraction was also markedly inhibited by catalase and EDTA. These results suggested that metal-catalyzed oxidation is involved in the GS inactivation by the small-molecular fraction of CSF. Dithiothreitol(DTT)was shown to inhibit almost completely the oxidative inactivation of GS by CSF. However, DTT inhibited only partially the oxidative inactivation of GS caused by small-molecular fraction of CSF. When large-molecular fraction of CSF was separated by anion-exchange HPLC chromatography, there was a peak of antioxidant activity inhibiting the small-molecular fraction-induced GS inactivation in the presence of DTT. The antioxidant activity was neutralized by monoclonal antibodies to transthyretin. Purified transthyretin was found to efficiently inhibit ascorbate/Cu2+-induced GS inactivation in the presence of DTT. Uric acid and glucose did not shoe any protective effect on the GS inactivation in the same condition. The above results suggest that metal-catalyzed oxidation occurs normally in human CSF, and the transthyretin may play an important role as a CSF antioxidant in protecting proteins from metal-catalyzed oxidation.

Key Words: Transthyretin; Cerebrospinal fluid; Antioxidant; Metal-catalyzed oxidation; Enzyme inactivation