Bergom Carmen, Paddock Cathy, Gao Cunji, Holyst Trudy, Newman Debra K, Newman Peter J
Blood Research Institute, BloodCenter of Wisconsin, 8727 Watertown Plank Road, Milwaukee, WI 53201, USA.
J Cell Sci. 2008 Apr 15;121(Pt 8):1235-42. doi: 10.1242/jcs.025163.
The Ig-ITIM family member PECAM-1 is expressed in vascular and endothelial cells, and its functions include suppression of mitochondria-dependent apoptosis. Previous studies have identified distinct PECAM-1 cytoplasmic domain splice variants at the mRNA, but not protein, level. Several relatively abundant mRNA isoforms lack exon 15 (Delta15) and would theoretically encode a protein with a truncated cytoplasmic domain and a unique C-terminal sequence. Using a novel rabbit polyclonal antibody that specifically recognizes Delta15 PECAM-1, we found that the Delta15 PECAM-1 isoform was expressed in human tissues, including brain, testes and ovary. This isoform was also expressed on the cell surface of human platelets, human umbilical vein endothelial cells (HUVECs) and the Jurkat T-cell leukemia, human erythroleukemia (HEL) and U937 histiocytic lymphoma cell lines. Furthermore, murine platelets and lung lysates demonstrated abundant amounts of exon-15-deficient PECAM-1. Functional studies revealed that Delta15 PECAM-1 retains both its homophilic binding capacity and its ability to signal by means of its immunoreceptor tyrosine-based inhibitory motif (ITIM) domains. Delta15 PECAM-1 was unable, however, to protect against apoptosis induced by overexpression of Bax or treatment with the chemotherapy agent etoposide. These studies suggest a novel role for the PECAM-1 C-terminus in cytoprotective signaling and highlight a need for further characterization of expression of PECAM-1 isoforms in normal and malignant tissues.
免疫球蛋白免疫受体酪氨酸抑制基序(Ig-ITIM)家族成员血小板内皮细胞黏附分子-1(PECAM-1)在血管和内皮细胞中表达,其功能包括抑制线粒体依赖性凋亡。先前的研究已在mRNA水平而非蛋白质水平鉴定出不同的PECAM-1胞质结构域剪接变体。几种相对丰富的mRNA异构体缺少外显子15(Delta15),理论上会编码一种具有截短胞质结构域和独特C端序列的蛋白质。使用一种能特异性识别Delta15 PECAM-1的新型兔多克隆抗体,我们发现Delta15 PECAM-1异构体在包括脑、睾丸和卵巢在内的人体组织中表达。这种异构体也在人血小板、人脐静脉内皮细胞(HUVECs)以及Jurkat T细胞白血病、人红白血病(HEL)和U937组织细胞淋巴瘤细胞系的细胞表面表达。此外,小鼠血小板和肺裂解物显示出大量缺乏外显子15的PECAM-1。功能研究表明,Delta15 PECAM-1保留了其嗜同性结合能力及其通过基于免疫受体酪氨酸的抑制基序(ITIM)结构域进行信号传导的能力。然而,Delta15 PECAM-1无法保护细胞免受Bax过表达或化疗药物依托泊苷处理诱导的凋亡。这些研究表明PECAM-1 C端在细胞保护信号传导中具有新作用,并强调需要进一步表征PECAM-1异构体在正常和恶性组织中的表达情况。