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J. Biochem, 2004, Vol. 136, No. 4 457-462
© 2004 The Japanese Biochemical Society

BIOCHEMISTRY

Cytotoxic and Membrane Perturbation Effects of a Novel Amyloid Forming Model Peptide Poly(Leucine-Glutamic Acid)

Rajadas Jayakumar*, Jayaraman Murali, Deenadayalan Koteeswari and Kannayiram Gomathi

Bio-organic and Neurochemistry Division, Central Leather Research Institute, Adyar, Chennai, 600 020, India

In the present study we have elucidated the toxicity of a novel amyloid forming model peptide, Poly (leucine-glutamic acid). The toxicity of the fibrils prepared from this peptide was analyzed in peripheral blood lymphocytes (PBL). The MTT reduction assay revealed that the viability of PBL decreases significantly upon treatment with Poly(leucine-glutamic acid) (Poly [LE]). Enhanced DCFH-DA fluorescence in treated cells suggests that peptide toxicity is probably mediated by the formation of free radicals. In vivo and in vitro biochemical studies indicated that Poly [LE] inactivates the antioxidant system of cells. Perturbation of Poly [LE] in a membrane lipid environment was assessed by circular dichroism (CD) using phosphotidyl choline-cholesterol bilayers. The CD results revealed that LE enhances its beta sheet content in a bilayer environment. Sequestration of Poly [LE] in lipid rafts demonstrates that it has a binding cleft similar to Aß in lymphocyte raft domains. Nuclear membrane binding studies showed that Poly [LE] binds to nuclear membranes and may cause genotoxicity.

* To whom correspondence should be addressed. Phone: +91-44-24911386 (Ext. 324), Fax: +91-44-24911589, E-mail: karkuvi77{at}hotmail.com


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