Team:Arizona State/Magainin

From 2012.igem.org

Revision as of 06:12, 3 October 2012 by Napatel (Talk | contribs)

Magainin-Split Beta-Galactosidase Biosensor

Overview

Current methods of water-borne pathogen detection require expensive equipment that often isn't modular, high throughput, or accessible to developing countries. One of the projects this year's ASU iGEM undertook was building a split-enzyme engineered fusion protein consisting of the Magainin peptide and the split alpha and omega fragments of beta-galactosidase.

Magainin

Magainins are a class of proteins found in the skin of Xenopus laevis[http://en.wikipedia.org/wiki/Xenopus_laevis] - the African clawed frog - that contain antimicrobial properties. Both Magainin 1 and 2 are closely related peptides of 23 amino acids each and only differ by two amino acid substitutions. These two peptides have broad spectrum antimicrobial activity, including gram-positive and gram-negative bacteria, viruses, protozoa, yeasts and fungi, and are hemolytic and cytotoxic to cancer cells. The mechanism for antimicrobial activity in Magainin 1 and 2 involves the disruption the plasma membranes of target cells via electrostatic interactions between the negatively charged phospholipid bilayer and the positively-charged amino-terminus of the Magainin peptide.

The design of the biosensor took advantage of the electrostatic interaction between the Magainin-1 peptide and the lipid bilayer of bacteria to design a bacterial biosensor that confers a signal based on the binding of multiple Magainin peptides to a bacterial membrane (Figure A).

Biosensor Design Concept

Figure A depicts the Magainin-Linker-Split Beta-galactosidase biosensor design concept. In close proximity, the two subunits of beta-galactosidase, Alpha and Omega, interact to form a complete, fully-functional unit. Because Magainin peptides congregate and form a toroidal pore when binding to the membranes of bacteria, the close proximity of the linked split beta-galactosidase units can theoretically be achieved.

Progress Map