1.C.28 The Bacteriocin AS-48 Cyclic Polypeptide (Bacteriocin AS-48) Family

Many organisms synthesize proteins (or peptides) which are degraded to relatively small hydrophobic or amphipathic, bioactive peptides. These peptides exhibit antibiotic, fungicidal, virucidal, hemolytic and/or tumoricidal activities by interacting with membranes and forming transmembrane channels that allow the free flow of electrolytes, metabolites and water across the phospholipid bilayers. Most of these peptides appear to function in biological warfare. There are many designations given to these bioactive peptides. They include the magainins, cecropins, melittins, defensins, bacteriocidins, etc. The proteins in each family within this functional superfamily are homologous, but they exhibit little or no significant sequence similarity with members of the other families. Thus, each family may have evolved independently. However, certain common structural features observed between members of distinct families suggest that at least some of these families share a common ancestry.

The generalized transport reaction catalyzed by channel-forming amphipathic peptides is:

small solutes, electrolytes and water (in) small solutes, electrolytes and water (out).

Bacteriocins are bacterially produced peptide antibiotics with the ability to kill a limited range of bacteria, usually but not always those that are closely related to the producer bacterium. Many of them exhibit structural features typical of members of the eukaryotic channel-forming amphipathic peptides. That is, they are usually synthesized as small precursor proteins or peptides which are processed with proteolytic elimination of their N-terminal leader sequences, and the resultant mature peptides form one, two or more putative amphipathic transmembrane α-helical spanners (TMSs). For those with two TMSs, a characteristic hinge region that separates the two putative transmembrane segments is usually observed. A similar structural arrangement occurs in the two-TMS Cecropin A proteins (TC #1.C.17).

Many bacteriocins are encoded in operons that also encode an immunity protein and an ABC transport system (TC #3.A.1) with a protease domain at the N-terminus. The ABC systems export the bacteriocins while the protease domains cleave the N-terminal leader sequence. A few bacteriocins are exported by the type II general secretory pathway rather than by ABC-type export systems. In some cases, expression of the bacteriocin-encoding operon is induced by a bacteriocin-like peptide which acts in conjunction with a two-component sensor kinase-response regulator to effect induction.

Many bacteriocins have been identified in addition to those tabulated in the TC system, but those listed are among the best characterized, with respect to evidence for channel formation in target bacterial membranes. Class III and IV bacteriocins (Klaenhammer, 1993) are large heat-labile proteins that function by mechanisms unrelated to those of the bacteriocins listed here.

Bacteriocin AS-48 is a 70 residue cyclic polypeptide derived from a 105 aa precursor encoded by the bacAgene on the pMB2 plasmid of Enterococcus faecalis. The protein consists of a globular arrangement of five α-helices. These helices enclose a compact hydrophobic core. This arrangement resembles that observed for NK lysin of the Amoebapore family (TC #1.C.35). The head-to-tail union lies in the middle of helix 5, and this union promotes stability of the 3-D structure. The cyclic polypeptide forms channels in membranes, accounting for its dissipation of the proton motive force and cell toxicity. Helix 4, which bears positive charges, has been suggested to participate directly in channel formation.