Characterization of the Proprotein Convertase-Mediated Processing of Peroxidasin and Peroxidasin-like Protein

Peroxidasin (PXDN) and peroxidasin-like protein (PXDNL) are members of the peroxidase-cyclooxygenase superfamily. PXDN functions in basement membrane synthesis by forming collagen IV crosslinks, while the function of PXDNL remains practically unknown. In this work, we characterized the post-translational proteolytic processing of PXDN and PXDNL. Using a novel knock-in mouse model, we demonstrate that the proteolytic cleavage of PXDN occurs in vivo. With the help of furin-specific siRNA we also demonstrate that the proprotein-convertase, furin participates in the proteolytic processing of PXDN. Furthermore, we demonstrate that only the proteolytically processed PXDN integrates into the extracellular matrix, highlighting the importance of the proteolysis step in PXDN’s collagen IV-crosslinking activity. We also provide multiple lines of evidence for the importance of peroxidase activity in the proteolytic processing of PXDN. Finally, we show that PXDNL does not undergo proteolytic processing, despite containing sequence elements efficiently recognized by proprotein convertases. Collectively, our observations suggest a previously unknown protein quality control during PXDN synthesis and the importance of the peroxidase activity of PXDN in this process.

Epithelial HVEM promotes basement membrane synthesis and intraepithelial T cell survival and migration

SummaryIntraepithelial T cells (IET) provide continuous surveillance of the intestinal epithelium, but little was known about how epithelial-derived signals regulate the IET population. We show that epithelial expression of the herpes virus entry mediator (HVEM), a member of the TNF receptor superfamily (TNFRSF), maintained the survival of small intestine IET, especially innate-like TCRαβ+ cells lacking CD4 and CD8β. Patrolling movement of all CD8α+ IET also was impaired in the absence of HVEM. HVEM-deficient epithelial cells exhibited downregulation of synthesis of basement membrane components, including collagen IV. Collagen IV supported IET survival in vitro via interactions with β1 integrins expressed by the IET; absence of β1 integrins decreased some IET subsets. Therefore, these data define a circuit whereby epithelial cells regulate intestine resident T lymphocyte populations through basement membrane synthesis.

The prognostic significance of type IV collagen expression in colorectal carcinomas

BACKGROUND: Breakdown of basement membrane is believed to be an essential step for tumor invasion and metastasis. The interaction between tumor cells and extracellular matrix can also result in induction of basement membrane synthesis by tumor and stromal cells. The aim of this study was to investigate distribution patterns of type IV collagen expression at the basement membrane in the tumor tissue of colorectal carcinomas by immunohistochemical staining. The degree of expression of type IV collagen correlated with classical clinicopathologic prognostic factors and their potential relationship with patients' prognosis. PATIENTS AND METHODS: This study included 40 patients who underwent curative resection of colorectal cancer at the Department of Surgery in the General Hospital Senta with complete follow-up for 5 years or until death. The identification of basement membrane was performed immunohistochemically using monoclonal antibodies to collagen IV. The basement membrane synthesis at the invasive front of colorectal cancer was semiquantitatively assessed as mild, moderate or severe. RESULTS: The deposition of basement membrane type of collagen IV had a statistically significant correlation with the stage of disease and histological grade of tumor with tendency of lesser synthesis of basement membrane in the advanced stage of disease and poorer histological grade of tumor. There were no significant correlations between intratumoral basement membrane synthesis and sex and age of patients, and localization and histological type of tumor. Five-year survival rates in patients with mild synthesis of basement membrane in colorectal tumor were significantly lower than in patients with severe expression of basement membrane. The basement membrane synthesis in colorectal cancer was a prognostic factor with significance behind stage of disease and histological grade of tumor by univariate Cox hazard-model. Multivariate Cox analysis verified that explicit synthesis of basement membrane showed a trend toward better prognosis, but it was not statistically significant. CONCLUSION Immunohistochemical staining for type IV collagen is an important additional technique with prognostic value.