Staphylococcin 1580, a bacteriocin produced by Staphylococcus epidermidis 1580, is bactericidal to sensitive cells of many gram-positive bacteria and stable staphylococcal L-forms. The bacteriocin inhibited simultaneously the syntheses of deoxyribonucleic acid, ribonucleic acid, and protein, and caused neither degradation of deoxyribonucleic acid nor induction of phages in lysogenic, sensitive cells. After 1 hr of treatment, extensive degradation of ribonucleic acid occurred, which was accompanied by leakage of ultraviolet-absorbing material out of the cell. The incorporation of glucose in acid-precipitable and glycogenlike material was inhibited. Furthermore, the staphylococcin inhibited the transport of glucose, glutamic acid, rubidium ions, and o-nitrophenyl-beta-galactoside. The uptake of oxygen was only gradually affected, but the intracellular adenosine triphosphate level fell rapidly to 15% of the control value. The motility of sensitive Bacillus subtilis cells was markedly reduced on treatment. Staphylococcin 1580 exhibited no phospholipase activity. The phenomena are interpreted as resulting from an altered conformation and composition of the membrane, from an inhibition of transport through the membrane, or from a combination of these effects.