Studies on Prolidase Deficiency - Effect of Iminodipeptides on Collagen Metabolism

脯氨酸酶缺乏症的研究——亚氨基二肽对胶原蛋白代谢的影响

• 批准号: 62570454
• 负责人: ARATA Jiro
• 金额: $ 1.09万
• 依托单位: Okayama University Medical School (1988)Kochi Medical School (1987)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570454/
• 关键词: Prolidase deficiency; Prolidase isozyme; Fibroblast; Wound healing; Mouse epidermis; 基質特異性; 2価イオン; アイソザイム; 症状発現; 遊離アミノ酸; プロリダーゼ; プロリールハイドロキシレース

1) The prolidase(PD) activities of cultured skin fibroblasts derived from prolidase deficient sisters, the elder with typical clinical manifestations and the younger with only slight clinical manifestations were examined biochemically. 2) Isozymes of prolidase(PD-I,II) from cultured human skin fibroblasts were separated by DESE-5PW column HPLC. 3) PD activities in rat skin during wound healing were studied. 4) PD activities in an extract of newborn mouse epidermis were examined. Results 1) PD activities against several substrates other than Gly-Pro were present to some degree in both sisters. There were no detectable differences in PD activity between symptom(+) patients and the symptom(-) sister with respect to the effect of divalent cations and the substrate specificity. 2) In control, activities of PD-1 were decreased in the descending order of activities against Gly-Pro, Ala-Pro, Met-Pro. However, activities of PD-II were decreased in the descending order of activities against Met- … More Pro, Gly-Pro, Ala-Pro. In prolidase deficient sisters, the activities of PD-I could not be detected, while the activities of PD-II were present. No remarkable differences were seen between the substrate specificities of the PD-II from symptom(+) patient and symptom(-) sister. 3) PD activities in rat skin adjacent to the wound were decreased at day 1 and gradually increased to about three times those of controls at day 14. 4) PD activities were detected in newdorn mouse epidermis. Mn^<2+> had a potent activating effect. Discussion 1) In this study, we estimated PD activity in cultured skin fibroblasts derived from a symptom(+) patient, her symptom(-) sister and controls. PD activities against substrates other than Gly-Pro were present to some degree in both patients in crude fibroblas extract. This indicates that PD was not absent but altered. 2) Recently, two forms of PD have been separated from human cells and various tissues. In our patients, activity of PD-I could not be determined in both sisters,while activity of PD-II present. The studies of substrate specificity using partially purified PD-II showed on differences between symptom(+) patient and symptom(-) sister. 3) The activities of PD adjacent to the wound were elevated at 14 days after injury. The local supply of amino acids produced by increased PD activity may compensate to some extent for amino acids used for synthesis of new collagen in wound healing. The relation of the dysfunction of the PD to the pathogenesis of the wound healing is unclear. However, insufficient recycling of endogenous free proline might cause delay in wound healing. 4) The existence of PD activities in the epidermis may suggest a possible involvement of PD in epitheilialization. In these contexts effect of iminodipeptides on collagen metabolism will be investigated. Less

1)用生化方法检测酪氨酸酶缺陷姐妹、临床表现典型的老年人和临床表现轻微的年轻人皮肤成纤维细胞的脯氨酸酶(PD)活性。2)用DESE-5PW色谱柱分离培养的人皮肤成纤维细胞中的丙氨酸氨基转移酶同工酶(PD-I、PD-II)。3)观察创面愈合过程中大鼠皮肤中PD活性的变化。4)对新生小鼠表皮提取液进行了PD活性测定。结果1)除Gly-Pro外，两姐妹均存在不同程度的PD活性。在二价阳离子的影响和底物特异性方面，症状(+)患者和症状(-)姐妹患者之间的PD活性没有明显差异。2)对照组PD-1活性由高到低依次为Gly-Pro、Ala-Pro、Met-Pro。而PD-II的活性按抗Met-…活性的降序递减更多的Pro，Gly-Pro，Ala-Pro。在脯氨酸酶缺乏的姐妹中，检测不到PD-I的活性，但存在PD-II的活性。症状(+)患者的PD-II底物特异性与症状(-)姐妹的PD-II的底物特异性无显著差异。3)大鼠创面附近皮肤的PD活性在第1天开始下降，第14天逐渐增加到对照组的3倍左右。4)新生小鼠皮肤中检测到PD活性。Mn~(2+)有较强的激活作用。讨论1)在这项研究中，我们估计了来自症状(+)患者、她的症状(-)姐妹和对照的培养的皮肤成纤维细胞中PD的活性。在两名患者的粗纤维细胞提取液中，均存在不同程度的针对甘氨酸-Pro以外的底物的PD活性。这表明帕金森病并没有消失，而是发生了变化。2)最近，从人体细胞和各种组织中分离出了两种形式的PD。在我们的患者中，两姐妹都不能测定PD-I的活性，但存在PD-II的活性。用部分纯化的PD-II对底物特异性的研究表明，症状(+)患者和症状(-)姐妹患者之间存在差异。(3)伤后14天，创面附近PD活性升高。PD活性增加所产生的局部氨基酸供应可能在一定程度上补偿了在伤口愈合过程中用于合成新胶原的氨基酸。PD功能障碍与创面愈合机制的关系尚不清楚。然而，内源性游离脯氨酸循环不足可能导致伤口愈合延迟。4)表皮中PD活性的存在可能提示PD可能参与了上皮化。在这些背景下，将研究亚氨基二肽对胶原代谢的影响。较少

项目成果

Oono,Takashi.: Journal of Dermatology. 15. 212-219 (1988)

大野隆：皮肤病学杂志。

Takashi Oono;Jiro Arata: The Journal of Dermatology. 115(3). (1988)

Takashi Oono；Jiro Arata：皮肤病学杂志。