Studies on the formation mechanism and functions of the covalently bound complex of hyaluronan with SHAP, the functional molecular entity of hyaluronan

透明质酸与透明质酸功能分子实体SHAP共价结合复合物的形成机制和功能研究

• 批准号: 17370041
• 负责人: KIMATA Koji
• 金额: $ 9.96万
• 依托单位: Aichi Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17370041/
• 关键词: hyaluronan; SHAP; inflammation; inter-α-trypsin inhibitor; complex-forming enzyme factor; hepatitis; TSG-6; compliment; インターα-トリプシンインピビター; ビクニン; サイトカイン; インタ-α-トリプシンインヒビター; factor H-related protein 1; TSG6

The covalently bound complex formed from inter-α-trypsin inhibitor (ITI) and hyaluronan (HA) by transesterification reaction, the SHAP (ITI heavy chain)-HA complex was investigated with regard to its physiological functions and serum enz; yme factors involved in the reaction. The candidate protein obtained from one of the library of serum fractions by the beforehand establisehd purification methods using a series of affinity columns was identified FHR-1 (factor H-related protein-1). However, the medium fraction of FHR-1 cDNA-transfected hepatocarcinoma HLF cells did not show any of the activity. Meanwhile, we found that the medium of the transfectants with cDNA of TSG-6 (TNFα-stimulated gene 6 product) had the dramatically increased activity that was yet retained after removal of TSG-6 by passing through the antibody affinity column, suggesting that the enzyme factors could not be TSG-6 itself and be induced with TSG-6 in the cells. Thus, we are now in the processes to the purification … More and identification of the factors. We previously developed the bikunin (ITI short chain)-knockout mouse system in order to investigate in vivo functions of the SHAP-HA complex where the complex was not formed. We found that the knockout mouse was resistant to the induction of acute hepatitis by the administration of D-galactosamine/E.coli lipopolysaccharide (LPS) in which the CD44-HA interaction-dependent attachment of neutrophils to live capillary sinusoidal cell surfaces was involved in response to the LPS stimuli. We then found using culthred leukocytes that the SHAP-HA complex activates the CD44-HA interaction-dependent cell adhesion more than 100 times, which is a decisive evidence for the involvement of the SHAP-HA complex in inflammatory cell response. We also found the good relationship between the SHAP-HA complex levels and the disease progression in the serum samples from patients of liver diseases, articular cartilage injury, gestoses, ovarian cancer etc, suggesting functional significances of the SHAP-HA complex in those diseases and the high reliability of the complex as a disease marker. Less

α-胰蛋白酶抑制剂(ITI)与透明质酸(HA)通过酯交换反应形成共价复合物SHAP(ITI重链)-HA复合物的生理功能及参与血清酶因子的研究；通过预先建立的纯化方法，使用一系列亲和柱从血清级分库中获得的候选蛋白质被鉴定为FHR-1（因子H相关）。然而，FHR-1 cDNA 转染的肝癌 HLF 细胞的培养基部分没有表现出任何活性。同时，我们发现转染子的培养基中含有 TSG-6（TNFα 刺激的基因 6）。产品）的活性显着增加，但在通过抗体亲和柱去除 TSG-6 后仍保留，表明酶因子不可能是 TSG-6 本身，并且在细胞中被 TSG-6 诱导。因此，我们现在正处于纯化和鉴定因子的过程中。我们之前开发了 bikunin（ITI 短链）敲除小鼠系统，以研究 SHAP-HA 复合物的体内功能。我们发现，敲除小鼠对 D-半乳糖胺/大肠杆菌脂多糖 (LPS) 诱导的急性肝炎具有抵抗力，其中 CD44-HA 相互作用依赖性附着。然后，我们使用培养的白细胞发现，SHAP-HA 复合物激活 CD44-HA 相互作用依赖性细胞粘附超过 100 次，这是对 LPS 刺激的反应。我们还发现肝病、关节病患者血清样本中 SHAP-HA 复合物水平与疾病进展之间存在良好关系。软骨损伤、妊娠、卵巢癌等，表明 SHAP-HA 复合物在这些疾病中的功能意义以及该复合物作为疾病标志物的高可靠性。

项目成果

Recognition of sulfation pattern of chondroitin sulfate by uronosyl 2-O-sulfotransferase

• DOI: 10.1074/jbc.m508816200
• 发表时间: 2005-11-25
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Ohtake, S;Kimata, K;Habuchi, O
• 通讯作者: Habuchi, O

Role of serum-derived hyaluronan-associated protein-hyaluronan complex in ovarian cancer.

• DOI: 10.3892/or.19.5.1245
• 发表时间: 2008-05
• 期刊: Oncology reports
• 影响因子: 4.2
• 作者: Yukihiko Obayashi;H. Yabushita;Kouhei Kanyama;M. Noguchi;Lisheng Zhuo;K. Kimata;A. Wakatsuki

軟骨部腫瘍-細胞外マトリックスの役割とその制御-

软骨肿瘤-细胞外基质的作用及其调节-

• 发表时间: 2005
• 作者: 西田 佳弘;細野 幸三;内堀 充敏;田畑 出;卓麗 聖;木全 弘治;石黒 直樹
• 通讯作者: 石黒 直樹

Hyaluronan(HA) and serum-derived hyaluronan-associated protein(SHAP)-HA complex as predictive markers of cervical ripening in premature labor.

透明质酸（HA）和血清来源的透明质酸相关蛋白（SHAP）-HA复合物作为早产宫颈成熟的预测标志物。

• 发表时间: 2008
• 期刊: Connect Tissue Res. 49(2)
• 作者: Kishida T;Yabushita H;Wakatsuki A;Zhuo L. Kimata K.
• 通讯作者: Zhuo L. Kimata K.

Equivalent involvement of inter-alpha-trypsin inhibitor heavy chain isoforms in forming covalent complexes with hyaluronan.

间α-胰蛋白酶抑制剂重链同种型与乙酰透明质酸形成共价复合物的过程相同。

• 发表时间: 2008
• 期刊: Connect Tissue Res. 49(1)
• 作者: Zhu L;Zhuo L;Watanabe H;Kimata K.
• 通讯作者: Kimata K.