Electron microscopic studies on collagen fibrillogenesis.

胶原纤维生成的电子显微镜研究。

• 批准号: 61570009
• 负责人: KOBAYASHI KUNIHIKO
• 金额: $ 1.28万
• 依托单位: Department of Anatomy, Nagoya University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61570009/
• 关键词: Collagen; Fubrillogenesis; Electron microscopy; Intermolecular association; negative staining; molecular bundle (SLS); 4D-staggered linear polymer; テロペプチド; Segment-long-spacing

1. We previously found that collagen molecules associate at pH 4.5 to specifically form 4D-staggered linear polymers, which are assumed to be the first intermediates in in vitro collagen fibrillogenesis. In this study, we confirmed that the 4D-staggered aggregates (containing various degrees of polymerization) are incorporated into native-type D-periodic fibrils under the physiological conditions. We intended to isolate the oligomers of the different polymericity by gel filtration and to assign the role of each oligomer in collagen fibrillogenesis. The trials were not yet successful.2. Conditions for electron microscopic identification of the association form of the molecules (dimer) at the first step of collagen self-assembly were investigated and individual methods were optimized. We noticed the direct correlation between negatively stained banding patern of type I collagen molecular bundle (SLS) and the distribution of hydrophobic amino acids along the molecule. The correlation was confirmed with type I and type II collagens.3. Non-helical parts of the collagen molecule (telopeptide) were revealed to be involved in the 4D-staggered association at pH 4.5.4. Electron microscopic examination of the sample taken at an early stage of the in vitro fibrillogenesis showed the existance of D-periodic fibrils in addition to thin filaments (preliminaly obsevation).

1. 我们之前发现胶原蛋白分子在 pH 4.5 下缔合，特异性形成 4D 交错线性聚合物，这被认为是体外胶原纤维形成的第一个中间体。在这项研究中，我们证实了 4D 交错聚集体（包含不同程度的聚合）在生理条件下并入天然型 D 周期原纤维中。我们打算通过凝胶过滤分离不同聚合度的低聚物，并确定每种低聚物在胶原纤维形成中的作用。试验尚未成功。2．研究了胶原自组装第一步中分子（二聚体）缔合形式的电子显微镜鉴定条件，并优化了各个方法。我们注意到 I 型胶原分子束 (SLS) 的负染带型与疏水性氨基酸沿分子的分布之间存在直接相关性。与I型和II型胶原的相关性得到证实。 3．胶原蛋白分子（端肽）的非螺旋部分被发现参与 pH 4.5.4 下的 4D 交错缔合。对体外纤维形成早期阶段采集的样品进行电子显微镜检查，结果显示除了细丝之外还存在 D 周期原纤维（初步观察）。

项目成果

小林邦彦他: 細胞. 19. 379-383 (1987)

小林邦彦 (Kunihiko Kobayashi) 等人：细胞。19. 379-383 (1987)

KOBAYASHI,KUNIHIKO: Collagen and Related Research. 6. 195-201 (1986)

小林国彦：胶原蛋白及相关研究。

Kobayashi, K. et al.: "Correlation between negative staining pattern and hydrophobic residues of collagen." Journal of Electron Microscopy. 35. 272-275 (35)

Kobayashi, K. 等人：“负染色模式与胶原蛋白疏水残基之间的相关性。”

KOBAYASHI,KUNIHIKO: Journal of Electron Microscopy. 35(Suppl). 2703-2704 (1986)

小林国彦：电子显微镜杂志。

KOBAYASHI,KUNIHIKO: Journal of Ultrastructure and Molecular Structure Research.

小林国彦：超微结构和分子结构研究杂志。