Gnanasekar Munirathinam, Ramaswamy Kalyanasundaram
Department of Biomedical Sciences, College of Medicine, University of Illinois, Rockford, IL 61107, USA.
Parasitol Res. 2007 Nov;101(6):1533-40. doi: 10.1007/s00436-007-0671-z. Epub 2007 Aug 9.
Translationally controlled tumor protein (TCTP) is one of the most abundantly expressed proteins in the filarial parasites as well as in the other organisms. Several functions have been suggested for TCTP family of proteins ranging from calcium binding to histamine release function. However, its physiological function is still a mystery. Previous studies showed that the expression of TCTP is increased several-fold during oxidative stress. In the present work, we report the putative antioxidant function of Brugia malayi TCTP (BmTCTP). When tested in vitro, rBmTCTP could be reduced by a variety of reducing agents including thioredoxin. Such reduced form of rBmTCTP was able to protect DNA from oxidative damage, suggesting that BmTCTP may have an antioxidant function in the parasite. Sequence analysis of filarial TCTPs revealed that there are three cysteine amino acids located in the central portion of the protein. Subsequent targeted residue modification studies showed that these cysteine residues in rBmTCTP are critical for its antioxidant function. To determine the significance of this finding, rBmTCTP was overexpressed in vivo in Escherichia coli and subjected to oxidative stress. These studies showed that rBmTCTP significantly protected cells form oxidative damage. Taken together, these findings suggest that BmTCTP might be functioning as a non-classical antioxidant protein in the filarial parasites.
翻译调控肿瘤蛋白(TCTP)是丝虫寄生虫以及其他生物体中表达最为丰富的蛋白质之一。TCTP蛋白家族具有多种功能,从钙结合到组胺释放功能不等。然而,其生理功能仍是个谜。先前的研究表明,在氧化应激期间TCTP的表达会增加数倍。在本研究中,我们报道了马来布鲁线虫TCTP(BmTCTP)的假定抗氧化功能。在体外测试时,重组BmTCTP(rBmTCTP)可被包括硫氧还蛋白在内的多种还原剂还原。这种还原形式的rBmTCTP能够保护DNA免受氧化损伤,表明BmTCTP可能在寄生虫中具有抗氧化功能。丝虫TCTP的序列分析显示,在该蛋白的中央部分有三个半胱氨酸氨基酸。随后的靶向残基修饰研究表明,rBmTCTP中的这些半胱氨酸残基对其抗氧化功能至关重要。为了确定这一发现的意义,rBmTCTP在体内的大肠杆菌中过表达并遭受氧化应激。这些研究表明,rBmTCTP能显著保护细胞免受氧化损伤。综上所述,这些发现表明BmTCTP可能在丝虫寄生虫中作为一种非经典抗氧化蛋白发挥作用。