Studies on fibronectin-like protein of Japanese flounder

日本牙鲆纤连蛋白样蛋白的研究

• 批准号: 07660269
• 负责人: TOYOHARA Haruhiko
• 金额: $ 1.41万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660269/
• 关键词: collagenase; flounder; fibronectin; zymography; gelatin; protease; 血漿

Novel gelatinoltic activities were detected in Japanese flounder muscle, skin and serum by using gelatin-zymography. This result suggested the activities were possibly due to distinct enzymes. One of the activities were supposed to be derived from fibronectin-like protein based on the molecular weifht (>200kDa), sensitivity against proteolysis, wide tissue distribution, and affinity to gelatin and heparin.Fibronectin-like protein was purified by gelatin-affinity chromatography from Japanese flounder serum, which showed the gelatinolytic activity. The purified protein showed the immunological cross-reactivity with anti-chicken fibronectin antibody and the cell-spreading activity on the primary culture cells derived from Japanese flounder embryo and EPC cells.Other glatinolytic proteinases were demonstrated in various fish tissues. By the inhibitor effect, serine-type enzymes (70k-120k) and metallo-type enzyme (<100k) were identified. Similar activities were demonstrated in rat tissues, but the activities were remarkably weak compared with those of fishes, suggesting that the collagen would be metabolized at higher rate in fish tissues than in mammal tisuues.

采用明胶酶谱法检测了比目鱼肌肉、皮肤和血清中新的明胶酶活性。这一结果表明，活性可能是由于不同的酶。根据其分子量（> 200 kDa）、对蛋白质水解敏感、组织分布广、对明胶和肝素具有亲和性等特点，推测其活性之一来自纤连蛋白样蛋白。纯化的蛋白质与鸡纤连蛋白抗体具有免疫交叉反应性，并在原代培养的牙鲆胚胎细胞和EPC细胞上具有细胞铺展活性，在多种鱼组织中也发现了其他的明胶分解蛋白酶。根据抑制剂的作用，确定了丝氨酸型酶（70 k-120 k）和金属型酶（<100 k）。在大鼠组织中也表现出类似的活性，但活性明显弱于鱼类，表明胶原在鱼类组织中的代谢速率高于哺乳动物组织。

项目成果

K.Sato et al: "Involvement of type V collagen in softening of fish muscle during short-term chilled storage" J.Agric.Food Chem.(印刷中). (1997)

K.Sato 等人：“短期冷藏期间 V 型胶原蛋白参与鱼肌肉软化”J.Agric.Food Chem.（出版中）。

H.Toyohara et al.: "Elted of maturation on the Contents of free and bound forms of hydrory prolinl in Ayn muscle" Fisheries Sciouce. (in press). (1997)

H.Toyohara 等人：“艾因肌肉中游离和结合形式的氢脯氨酸含量的成熟”Fisheries Sciouce。