Interactions of melittin, a preprotein model, with detergents

Abstract

Bee venom melittin is a water-soluble tetramer of identical polypeptide chains. Each chain has 26 residues. The 20 N-terminal residues are hydrophobic and the 6 C-terminal residues are basic. Melittin integrates into natural and synthetic membranes and lyses a wide variety of cells. To understand how a water-soluble protein can spontaneously partition into a membrane, the interaction of melittin with micelles of deoxycholate (DOC), Brij 58 and sodium dodecyl sulfate (NaDodSO4) was studied. Circular dichroic spectra showed that NaDodSO4, an ionic detergent, and Brij 58, a nonionic detergent, caused similar major changes in the protein''s conformation. Gel filtration studies revealed that melittin forms mixed micelles with either Brij or DOC. The melittin-DOC mixed micelles have 2 mol of DOC/mol of melittin. Cross-linking studies with dimethyl suberimidate confirmed that the protein is a tetramer and showed that it becomes monomeric either in mixed micelles with Brij or DOC or in butanol. Despite this major structural change of melittin in the presence of an amphiphile, the covalently cross-linked form is as active in human erythrocyte lysis as the native protein.