Importance of the residue 190 on bactericidal activity of the bactericidal/permeability-increasing protein 5

Abstract

The bactericidal/permeability-increasing protein (BPI) with bactericidal and endotoxin-neutralizing activity is of considerable interest in clinical applications. However, the crucial residues responsible for the bactericidal activity of BPI remain elusive. In previous study, we identified the mutation of mBPI5 associated with the male infertility of mice. Here, the effects of Asp190Ala mutation on the antibacterial activity of mBPI5 have been determined. Substitution of Asp190 by alanine caused significant improvement in cytotoxic effect toward both E.coli J5 and P.aeruginosa. Liposome co-sedimentation assay showed that the ratio of Asp190Ala mutant binding to lipids increased by 8 folds. These results were well consistent with known fact that antibacterial activity of BPI is attributed to its high affinity for lipid moiety of lipopolysaccharides (LPS). The constructed structure of mBPI5 revealed that Asp190 was located close to 6 positively charged residues on the surface of N-terminal domain. When replacing Asp190 with alanine, salt linkages with Arg188 were broken, making the side chain of Arg188 be free to move and form tighter contacts with negatively charged LPS. These findings suggest that residue 190 combined with surrounding positively charged residues largely contribute to bactericidal and endotoxin-neutralizing activities of mBPI5.

Mammals' humoral immune proteins and peptides targeting the bacterial envelope: from natural protection to therapeutic applications against multidrug‐resistant Gram‐negatives

“…A ≈ 2 log unit reduction in multidrug‐resistant PA and AB viable cells was shown in vitro after 1 h with achievable doses of rBPI‐21, although KP remained mostly resistant (Weitz et al ., 2013 ). The Asp190Ala variant of BPI5 (murine BPI fold containing family B, member 5; also known as Bpifb5 or Lplunc5) showed a significant increase in LPS‐binding capacity and in in vitro antipseudomonal power (Wu et al ., 2016 ). Some synthetic constructs combining fragments of chemokines and functional residues of the BPI β‐sheet have shown interesting results: Betapep‐25 and its shorter dodecapeptide, SC‐4, showed a lethal dose 50 (LD 50 ) at nanomolar range for PA (Lockwood et al ., 2004 ; Dings, Haseman & Mayo, 2008 ; Dings et al ., 2013 ).…”