| Project/Area Number |
62560277
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
畜産化学
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| Research Institution | Obihiro University of Agriculture and Veterinary Medicine |
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Principal Investigator |
SHIMAZAKI Kei-ichi Obihiro Univ. of Agr. & Vet. Med. Asso. Prof., 畜産学部, 助教授 (10091547)
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| Co-Investigator(Kenkyū-buntansha) |
URASHIMA Tadasu Obihiro Univ. of Agr. & Vet. Med. Instructor, 畜産学部, 助手 (80185082)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1988: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1987: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | lactoferrrin / bovine milk protein / secondary structure / iron-binding / dye affinity / tryptic fragment / トリプシンフラグメント / 牛乳タンパク質 / 糖含量 / アミノ酸組成 / 糖鎖構造 / ホエータンパク質 / トリプシン分解 |
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| Research Abstract |
1. Molecular weight, sugar content and secondary structure of lactoferrin Molecular weight of lactoferrin was estimated to be 81,800+2,600 by SDSpolyacrylamide gel electrophoresis. Carbohydrate contents were 8,8 % for neutral sugar and 2.7 % for amino sugar. Helical content was 24 % calculated from CD spectrum.
2. Interacting properties of lactoferrin with blue dye Bovine lactoferrin showed strong interaction with immobilized Cibacron Blue F3GA column. It was suggested that such interaction has been due to the strong electrostatic forces.
3. Tryptic fragments of lactoferrin After digestion of lactoferrin by trypsin, fragments of which molecular weight was 52 KD and 27 KD were obtained. By the affinity chromatography with immobilized blue dye, the fragment of larger molecular weight showed the binding ability both to blue dye and iron. Onto the immobilized anti-lactoferrin serum column, the fragments of which molecular weights were from 20 to 40 KD, were adsorbed.
4. Estimation of composition and structure in carbohydrate moiety Carbohydrate moieties were liberated by hydrazinolysis and then were separated into neutral and acidic sugar moieties by ion exchange chromatography. They were estimated on their sugar compositions by gas chromatography. Immobilized endoblycosidases were also used to cleave the carbohydrate moieties in lactoferrin. Enzyme lectin assay was done in order to estimate the composition and the structure of the carbohydrate chains.
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