Molecular design of LPS-binding peptides.
Lipopolysaccharide (LPS), a major constituent of the outer membrane of Gram-negative bacteria, is highly toxic and can cause sepsis or septic shock. Therefore, detection of LPS and the ability to neutralize its toxicity is important. We previously obtained a strong LPS-binding peptide, Li5-001, using the phage display method (Matsumoto et al., 2010. J. Microbiol. Methods. 82, 54-58). We modified the sequence the amino acid sequence of this peptide (KNYSSSISSIHAC), by replacing and deleting amino acids to obtain higher LPS-binding affinity and greater resistance to protease digestion. Consequently we obtained a dodecapeptide, Li5-025 (K'YSSSISSIRAC', K' and C' are D-forms of K and C, respectively) which showed a high affinity for LPS, approximately 1000 folds higher affinity than Li5-001 and Kd value of 0.01 nM. By replacing both N- and C-terminal amino acids from L-type to D-type, the peptide was rendered resistant to protease digestion without altering its overall binding capacity.
Suzuki
Masatsugu Matt
MM
Peptide Door Co., Ltd., Sangyosozo-kan #204, Shimonojo 936-14, Takasaki, Gunma 370-0854, Japan.
Matsumoto
Megumi
M
Yamamoto
Akihiko
A
Ochiai
Masaki
M
Horiuchi
Yoshinobu
Y
Niwa
Makoto
M
Omi
Hiroyuki
H
Kobayashi
Tomomi
T
Takagi
Takashi
T
D019943:Amino Acid Substitution / Q000235:genetics; D008070:Lipopolysaccharides / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding
D008070:Lipopolysaccharides; D019151:Peptide Library; D010455:Peptides; D010447:Peptide Hydrolases