Inhibition of the interaction of urokinase-type plasminogen activator (uPA) with its receptor (uPAR) by synthetic peptides
Inhibition of the interaction of urokinase-type plasminogen activator (uPA) with its receptor (uPAR) by synthetic peptides
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DOI：
10.1515/bchm.1997.378.3-4.231
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发表时间：
1997-03-01
影响因子：
3.7
通讯作者：
Magdolen, V
中科院分区：
文献类型：
--
作者：
Burgle, M;Koppitz, M;Magdolen, V
丝氨酸蛋白酶尿激酶型纤溶酶原激活物（uPA）通过与其特异性受体（uPAR，CD 87）相互作用而聚集到细胞表面是肿瘤细胞侵袭和转移的重要步骤。通过定量流式细胞荧光分析（FACS）评估源自uPA的uPAR结合区（包含uPA的氨基酸16-32; uPA（16-32））的合成肽抑制荧光标记的uPA与人早幼粒细胞U937细胞上的uPAR结合的能力，并与已知干扰uPA/uPAR相互作用的其他合成肽的抑制能力进行比较。约3000倍摩尔过量的uPA（16-32）导致50%的pro-uPA与细胞表面相关uPAR结合的抑制。采用固相uPA-配体结合试验，采用包被在微量滴定板上的重组可溶性uPAR，确定了野生型uPA的最小结合区域。线性肽uPA（19-31）和其更稳定的二硫键桥环形式（cyclo（19，31）uPA（19-31））显示uPAR结合活性，而其它肽如uPA（18-30）、uPA（20-32）或uPA（20-30）不与uPAR反应。其中某些氨基酸缺失和/或被其它氨基酸取代的环（19，31）uPA（19-31）的环肽衍生物以及uPAR衍生的野生型肽也不抑制uPA/uPAR相互作用。因此，本研究确定了环（19，31）uPA（19-31）作为开发uPA-肽类似物以阻断uPA/uPAR相互作用的潜在先导结构。
Focusing of the serine protease urokinase-type plasminogen activator (uPA) to the cell surface via interaction with its specific receptor (uPAR, CD87) is an important step for tumor cell invasion and metastasis. The ability of a synthetic peptide derived from the uPAR-binding region of uPA (comprising amino acids 16-32 of uPA; uPA(16-32)) to inhibit binding of fluorescently labeled uPA to uPAR on human promyeloid U937 cells was assessed by quantitative flow cytofluorometric analysis (FACS) and compared to the inhibitory capacities of other synthetic peptides known to interfere with uPA/uPAR-interaction. An about 3000-fold molar excess of uPA(16-32) result in 50% inhibition of pro-uPA binding to cell surface-associated uPAR. Using a solid-phase uPA-ligand binding assay employing recombinant soluble uPAR coated to microtiter plates, the minimal binding region of wildtype uPA was determined, The linear peptide uPA(19-31) and its more stable disulfide-bridged cyclic form (cyclo(19,31) uPA(19-31)) displayed uPAR-binding activity whereas other peptides such as uPA(18-30), uPA(20-32) or uPA(20-30) did not react with uPAR. Cyclic peptide derivatives of cyclo(19,31) uPA(19-31) in which certain amino acids were deleted and/or replaced by other amino acids as well as uPAR-derived wild-type peptides did also not inhibit uPA/uPAR-interaction. Therefore, the present investigations identified cyclo(19,31) uPA(19-31) as a potential lead structure for the development of uPA-peptide analogues to block uPA/uPAR-interaction.