Fatty acids covalently bound to a-hemolysin of Escherichia coli are involved in

VANESA HERLAX; LAURA BAKÁS

BIOCHEMISTRY

Año: 2006 p. 1 - 1

0006-2960

a-Hemolysin (HlyA) is a pore-forming toxin secreted by pathogenic strains of Escherichia coli.  The toxin is synthesized as a protoxin, ProHlyA, which is matured in the cytosol to the active form by acylation at two internal lysines, K563 and K689 (HlyA). It is widely known that the presence of fatty acids is crucial for the hemolytic and cytotoxic effects of the toxin. However, no detailed physico-chemical characterization of the structural changes produced by fatty acids in the soluble protein prior to membrane binding has been carried out to date. The effects of chemical denaturants, the ANS binding parameters (Kd and n) and the sensitivity to proteases were compared between the acylated and unacylated protein forms HlyA and ProHlyA. Our results are consistent with a molten globular form of the acylated protein. Moreover, as molten globule proteins are intrinsically disordered proteins, using disorder prediction analyses, we show that HlyA contains nine regions composed of 10–30 natively disordered amino acids. We propose that this conformation induced by covalently bound fatty acids might provide HlyA with the ability to bind to a variety of molecules during its action mechanism.