Previously, we revealed that a cationic antibacterial polypeptide of 11 kDa (CAP11), a member of the cathelicidins isolated from guinea pig neutrophils, exhibits not only potent antibacterial activity but also lipopolysaccharide (LPS)-neutralizing activity. In this study, to determine the biologically active regions of CAP11, we isolated or synthesized the partial peptides of CAP11 and evaluated their antibacterial and LPS-neutralizing activities. Although CAP11 has a unique homodimeric structure with a disulfide bridge, the biological activities of dimeric and monomeric forms of CAP11 were almost the same. Moreover, the G(1)-E(33) peptide of CAP11 showed the same activities as CAP11, whereas the C-terminal region (Y(34) to I(43)) possessed no biological activities. In addition, the three 18-mer peptides (G(1)-R(18), T(9)-K(26), and L(16)-E(33)) with overlapping sequences were synthesized, and their activities were determined. The three 18-mer peptides retained the antibacterial activities, and G(1)-R(18) was the most potent. In contrast, the LPS-neutralizing activities of these peptides were markedly reduced. Together, these observations indicate that the active region with antibacterial activity is localized at G(1) to R(18) of CAP11, whereas longer sequences (such as G(1) to E(33)) would be required for the expression of LPS-neutralizing activity. Furthermore, the C-terminal region (Y(34) to I(43)) and a disulfide bridge are not essential for the antibacterial and LPS-neutralizing activities of CAP11.