1987 Fiscal Year Final Research Report Summary
A hypothesis as to mechanism of perturbation in biomembrane due to endotoxin. Participation of lysosomal cationic proteins with an endotoxin binding capacity
| Project/Area Number |
60480167
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
細菌学
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| Research Institution | Ieate Medical University, School of Medicine |
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Principal Investigator |
YOSHIDA Masao Iwate Medical University, School of Medicine, Professor, 医学部, 教授 (50048229)
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| Co-Investigator(Kenkyū-buntansha) |
TSUNODA Nobuko Iwate Medical University, School of Medicine, Assistant, 医学部, 助手 (90128926)
INADA Katsuya Iwate Medical University, School of Medicine, Lecturer., 医学部, 講師 (80048446)
HIRATA Michimasa Iwate Medical University, School of Medicine,Lecturer., 医学部, 講師 (20048359)
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| Project Period (FY) |
1985 – 1987
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| Keywords | Endotoxin / Granulocytes / Cationic protein / Mode of action / Biomembrane / Endotoxin binding protein / Hemagglutination / LPS |
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| Research Abstract |
1) Fractionation of cationic proteins (CAP's); CAP's were fractionated from rebbit peritoneal granulocytes through the steps of 0.1 M citric acid extraction, ethanol precipitation and gel filtration by FPLC. Since CAP's were capable of binding with heparin, a purification using heparin Sepharose is being investigated.
2) CAP's agglutinated the erythrocytes sensitized with LPS, especially Re-LPS (CAP-HA); CAP-HA was inhibited by preincubation of CAP with LPS.
3) CAP bound with LPS, especially Re-LPS in water solution, and made increased turbidity; the increase of turbidity depended upon CAP-levels, although the binding took place at an optimal ratio of both CAP and LPS levels; Low ionic strength condition and pH 6-7 were optimal to CAP-HA; the binding was reversible. This seems to be involved in both ionic and hydrophobic bounds of CAP and LPS. CAP-HA is thought to result from the binding mentioned above.
4) CAP exhibited antibacterial activity against S and R form gram-negative bacteria as well as gram-positive bacteria, expecially against R from gram-negatives. The activity seems to be bacteriolytic.
5) CAP bound with heparin made increased trubidity, consequently the binding neutralized mutually the properties of CAP and heparin.
6) One CAP fraction of less than 10,000 M.W. had high anticoagulant activity compared to other properties.
7) It seems that CAP can promote the activity of LPS, which bound with cells such as erythrocytes, and membrane perturbation.
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