Department of Pharmacology, University of the Basque Country, UPV/EHU, Leioa, Bizkaia, Spain; Centro de Investigación Biomédica en Red de Salud Mental (CIBERSAM), Spain.
Sorbonne Université, Neuroscience Paris Seine - IBPS, Team Neurobiology of Psychiatric Disorders, Paris, France; CNRS UMR8246, Neuroscience Paris Seine - IBPS, Team Neurobiology of Psychiatric Disorders, Paris, France; Inserm U1130, Neuroscience Paris Seine - IBPS, Team Neurobiology of Psychiatric Disorders, Paris, France.
Neuroscience. 2021 Jul 15;467:91-109. doi: 10.1016/j.neuroscience.2021.05.017. Epub 2021 May 24.
Hevin is a matricellular glycoprotein that plays important roles in neural developmental processes such as neuronal migration, synaptogenesis and synaptic plasticity. In contrast to other matricellular proteins whose expression decreases when development is complete, hevin remains highly expressed, suggesting its involvement in adult brain function. In vitro studies have shown that hevin can have different post-translational modifications. However, the glycosylation pattern of hevin in the human brain remains unknown, as well as its relative distribution and localization. The present study provides the first thorough characterization of hevin protein expression by Western blot in postmortem adult human brain. Our results demonstrated two major specific immunoreactive bands for hevin: an intense band migrating around 130 kDa, and a band migrating around 100 kDa. Biochemical assays revealed that both hevin bands have a different glycosylation pattern. Subcellular fractionation showed greater expression in membrane-enriched fraction than in cytosolic preparation, and a higher expression in prefrontal cortex (PFC) compared to hippocampus (HIP), caudate nucleus (CAU) and cerebellum (CB). We confirmed that a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4) and matrixmetalloproteinase 3 (MMP-3) proteases digestion led to an intense double band with similar molecular weight to that described as SPARC-like fragment (SLF). Finally, hevin immunoreactivity was also detected in human astrocytoma, meningioma, cerebrospinal fluid and serum samples, but was absent from any blood cell type.
Hevin 是一种细胞外基质糖蛋白，在神经发育过程中发挥重要作用，如神经元迁移、突触形成和突触可塑性。与其他细胞外基质蛋白不同，当发育完成时，其表达水平降低，而 hevin 仍然高度表达，表明其参与了成年大脑的功能。体外研究表明，hevin 可以发生不同的翻译后修饰。然而，人类大脑中 hevin 的糖基化模式及其相对分布和定位仍然未知。本研究通过 Western blot 首次对死后成人脑中的 hevin 蛋白表达进行了全面表征。我们的结果表明，hevin 有两种主要的特异性免疫反应条带：一条强烈的条带迁移约 130 kDa，另一条迁移约 100 kDa。生化分析表明，两条 hevin 条带具有不同的糖基化模式。亚细胞分离显示，膜富集部分的表达高于胞质部分，且前额叶皮层（PFC）的表达高于海马（HIP）、尾状核（CAU）和小脑（CB）。我们证实，解整合素金属蛋白酶 3（ADAMTS3）和基质金属蛋白酶 3（MMP-3）蛋白酶消化导致一条强烈的双带，其分子量与 SPARC 样片段（SLF）相似。最后，hevin 免疫反应性也在人类星形细胞瘤、脑膜瘤、脑脊液和血清样本中检测到，但不存在于任何血细胞类型中。
