Novel therapeutic strategy in the treatment of infectious diseases by anti-microbial, endotoxin-neutralizing proteins.
| Project/Area Number |
10670267
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Bacteriology (including Mycology)
|
|---|
| Research Institution | Iwate Medical University |
|---|
Principal Investigator |
HIRATA Michimasa Iwate Medical University, Department of Bacteriology, Associate professor., 医学部, 助教授 (20048359)
|
|---|
| Project Period (FY) |
1998 – 2000
|
| Project Status |
Completed (Fiscal Year 2000)
|
| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥1,500,000 (Direct Cost: ¥1,500,000)
|
|---|
| Keywords | Lipopolysaccharide (LPS) / Endotoxin-binding protein / Endotoxinneutralization / CAP18 peptides / Antibacterial peptide / Innate immunity / Host defense mechanism / 難治性感染症 / エンドトキシン結合蛋白 / 合成ペプチド / エンドトキシン中和 / エンドトキシン結合ドメイン / 抗菌活性 |
|---|
| Research Abstract |
We have found 18kDa cationic anti-microbial proteins (CAP18) with LPS-neutralizing activity from rabbit and human neutrophils. The 27mer C-terminal fragment of human CAP 18 (27mer ; CAP18_<109-135>) has been identified as the anti-microbial and LPS-neutralizing domain. We investigated the effects of modification and/or substitution of amino acid residues of CAP18 peptides on their activities. Acetylation of N-terminal and amidation of C-terminal residues of the 27mer in-creased LPS-neutralizing and antimicrobial activities. Substitution of E_<119> and K_<128> of 27mer to L (27LL) or F (27FF) increased LPS binding activity (〜30-40 fold) and also blocked LPS-induced lethality in mice. Substitution of E_<119> to L_<119> increased LPS-binding activity (〜8 -fold), however, this peptide (27L) could not protect vs. endotoxin shock. Truncation of 9 amino acids from the N-terminus of 27mer (18mer : CAP18_<118-135>) resulted in a significant decrease in LPS-binding activity even though both terminuses are chemically modified. Increasing the hydrophobic amino acid residues (Leucin=L) in 18mer (18LL) increased LPS-binding activity (〜30-fold) and anti-microbial activity versus gram-positive bacteria such as MRSA (methicillin resistant staphylococcus aureus ; >3 -folds), however, 18LL could not protect vs. endotoxin shock in mice. Further replacement of three positions of 18LL by lysine (K) significantly increased LPS-binding activity (〜120-fold), antimicrobial activity vs. gram negative bacteria such as E.coli O157 : H7 and S.typhimurium, and protect mice from endotoxin shock.
From these results, the importance of the stability of alpha-helical structure, and the balance of hydrophobic/ hydrophilic (basic) amino acid residues in CAP18 peptide was suggested.
|
Report
(4 results)
Research Products
(30 results)
-
-
-
-
-
-
-
-
-
-
-
-
-
[Publications] Kirikae, T., Hirata, M., Yamasu, H., Kirikae, F., Tamura, H., Kayama, F., Nakatsuka, K., Yokochi, T., Nakano, M: "Protective effects of a human 18-kilodalton cationic antimicrobial protein (CAP18)-derived peptide against murine endotoxemia."Infect.Immun. 66. 1861-1868 (1998)
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Publications] Nagaoka, I., Hirata, M., Sugimoto, K., Tsutsumi-Ishii, Y., Someya, A., Saionji, K., Igari, J: "Evaluation of the expression of human CAP18 gene during neutrophil maturation in the bone marrow"J.Leukoc.Biol. 64. 845-852 (1998)
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Publications] Osaki, T., Omotezako, M., Nagayama, R., Hirata, M., Iwanaga, S., kasahara, J., Hattori, J., Ito, I., Sugiyama, H., Kawabata, S: "Horseshoe crab hemocyte- derived antimicrobial polypeptides, tachystatins, with sequence similarity to spider neurotoxins."J.Biol.Chem.. 274. 26172-26178 (1999)
Description
「研究成果報告書概要(欧文)」より
Related Report
-
-
[Publications] Swierzko, AS., Kirikae, T., Kirikae, F., Hirata, M., Cedzynski, M., Ziolkowski, A., Hirai, Y., Kusumoto, S., Yokochi, T., Nakano, M: "Biological activities of lipopolysaccharides of proteus spp. and their interactions with polymyxin B and an 18-kDa cationic antimicrobial protein (CAP18)-derived peptide"J.Med.Microbiol. 49. 127-138 (2000)
Description
「研究成果報告書概要(欧文)」より
Related Report
-
-
-
-
-
-
-
-
-
-
-
-
-
