The DNA repair factor ku80 binds and activates the adhesion receptor ELTD1/ADGRL4

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications Pub Date : 2025-04-08 DOI:10.1016/j.bbrc.2025.151785

Stefan Amisten , Melinda Rezeli , Mario Grossi , Paulina Bryl-Gorecka , Anton Håkansson , Kristina Torngren , György Marko-Varga , David Erlinge , Björn Olde

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引用次数: 0

Abstract

We investigated the mRNA expression of G protein-coupled receptors (GPCRs) in cells from four human vascular beds: umbilical vein (HUVEC), microvasculature (MVEC), aorta (HAEC), and coronary artery (CAEC). Our study revealed that the orphan receptor ELTD1 (ADGRL4) was the most abundantly expressed GPCR mRNA in all four EC types. When recombinantly expressed in U87 cells, ELTD1 receptors activated canonical GPCR pathways, particularly the Gq pathway. Conditioned medium from U87 cells also activated ELTD1 in HEK293 cells, supporting the existence of an autocrine ELTD1-activating factor. Using affinity capture and mass spectrometry in combination with the label free xCelligence system, we identified the proteins ku80 and erythrocyte beta spectrin (SPTB) as ligands to ELTD1.

Ku80 demonstrated higher potency in activating ELTD1 than SPTB, thus leading us to focus on this protein. INCA-X, a functional antibody targeting the ku80/ku70 complex, and ELTD1 siRNA impaired endothelial tube formation in a similar manner, suggesting a common pathway. In a study cohort of myocardial infarction patients, high plasma levels of the extracellular domain of ELTD1 correlated (P < 0.05) with high (>2.9) cardiovascular flow reserve, thus indicating an association between ELTD1 and active angiogenesis and revascularization.

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DNA修复因子ku80结合并激活粘附受体ELTD1/ADGRL4

我们研究了四种人类血管床细胞中G蛋白偶联受体（gpcr）的mRNA表达：脐静脉（HUVEC）、微血管（MVEC）、主动脉（HAEC）和冠状动脉（CAEC）。我们的研究显示孤儿受体ELTD1 （ADGRL4）是所有四种EC类型中表达最丰富的GPCR mRNA。当在U87细胞中重组表达时，ELTD1受体激活了典型的GPCR途径，特别是Gq途径。来自U87细胞的条件培养基也激活了HEK293细胞中的ELTD1，支持自分泌ELTD1激活因子的存在。利用亲和捕获和质谱结合无标签xCelligence系统，我们鉴定出ku80蛋白和红细胞β谱蛋白（SPTB）是ELTD1的配体。Ku80在激活ELTD1方面表现出比SPTB更高的效力，因此我们将重点放在该蛋白上。靶向ku80/ku70复合物的功能性抗体INCA-X和ELTD1 siRNA以类似的方式破坏内皮管的形成，提示有共同的途径。在一项心肌梗死患者队列研究中，高血浆水平的ELTD1细胞外结构域与(P <；0.05)，心血管血流储备高（>2.9），因此表明ELTD1与活跃的血管生成和血运重建之间存在关联。

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来源期刊

Biochemical and biophysical research communications 生物-生化与分子生物学

CiteScore

6.10

自引率

0.00%

发文量

1400

审稿时长

14 days

期刊介绍： Biochemical and Biophysical Research Communications is the premier international journal devoted to the very rapid dissemination of timely and significant experimental results in diverse fields of biological research. The development of the "Breakthroughs and Views" section brings the minireview format to the journal, and issues often contain collections of special interest manuscripts. BBRC is published weekly (52 issues/year).Research Areas now include: Biochemistry; biophysics; cell biology; developmental biology; immunology ; molecular biology; neurobiology; plant biology and proteomics