Biological significance of Prolinerich antimicrobial peptide

富含脯氨酸抗菌肽的生物学意义

• 批准号: 08044228
• 负责人: KOHGO Yutaka
• 金额: $ 2.05万
• 依托单位: Asahikawa Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08044228/
• 关键词: antimicrobial peptide; PR-39; syndecan; metastasis; cytoskeletone; ras sinal; 内因性抗菌ペプチド

We clarified several important points about syndecans and PR-39 through colaborations and discussions with Dr.Gallo as shown below.1.We made polyclonal antibody against PR-39 afer immunization of rabbits using N-terminal 15 aminoacids oigopeptides.2.We observed the band at the size of approximately 5 kDa as expected size of PR-39 mature protein afer immunoprecipitation of human leukocytes protein using PR-39 polyclonal antibody. However, we have not cloned the protein which might be human counterpart of pig PR-39.3.PR-39 has a possible function which can bind to Src homology 3 domain, becaus PR-39 has five repeats of proline rich motif that has been reported to bind SH3 domain as like a Sos protein which can bind to Grb2 protein. Recombinant SH3 domain of Src gene can actually bind to PR-39 oligopeptides by the study of immunopregipitation.4.PR-39 gene transfection into human hepatoma cells showed induction of syndecan-1 expression, suppression of invasive activity and cell morphological change. These results might be due to the binding ability of PR-39 into SH3 domain.5.PR-39 transfectans excreted PR-39 protein into conditioning medium.6.PR-39 gene transfection into ras transformants also showed morphological change and decrease of proliferation rate.

通过与Gallo博士的合作和讨论，我们澄清了关于syndecans和PR-39的几个要点，如下所示。1.我们使用N端15个氨基酸寡肽免疫兔子后制备了针对PR-39的多克隆抗体。2.我们观察到使用 PR-39 多克隆抗体对人白细胞蛋白进行免疫沉淀后，条带大小约为 5 kDa，与 PR-39 成熟蛋白的预期大小相同。然而，我们还没有克隆出可能是猪 PR-39.3 的人类对应物的蛋白质。PR-39 可能具有与 Src 同源 3 结构域结合的功能，因为 PR-39 具有已报道的富含脯氨酸基序的五个重复结合SH3结构域，就像可以结合Grb2蛋白的Sos蛋白一样。免疫沉淀研究发现Src基因重组SH3结构域确实能与PR-39寡肽结合。4.PR-39基因转染人肝癌细胞后可诱导syndecan-1表达，抑制侵袭活性和细胞形态改变。这些结果可能是由于PR-39与SH3结构域的结合能力所致。5.PR-39转染子将PR-39蛋白分泌到条件培养基中。6.PR-39基因转染到ras转化子中也表现出形态变化和增殖减少速度。

项目成果

Matsumoto A: "Ruduced expression of syndecan-1 in human hepatocellular carcinoma with high metastatic potential." Int.J.Cancer. 74. 482-491 (1997)

Matsumoto A：“具有高转移潜力的人肝细胞癌中 syndecan-1 的表达减少。”

Matsumoto A: "Ruduced expression of syndecan-1 in human hepatocellular carcinoma with high metastatic potential." Int. J. Cancer. 74. 482-491 (1997)

Matsumoto A：“具有高转移潜力的人肝细胞癌中 syndecan-1 的表达减少。”