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KLK3/PSA and cathepsin D activate VEGF-C and VEGF-D.

ABSTRACT: Vascular endothelial growth factor-C (VEGF-C) acts primarily on endothelial cells, but also on non-vascular targets, for example in the CNS and immune system. Here we describe a novel, unique VEGF-C form in the human reproductive system produced via cleavage by kallikrein-related peptidase 3 (KLK3), aka prostate-specific antigen (PSA). KLK3 activated VEGF-C specifically and efficiently through cleavage at a novel N-terminal site. We detected VEGF-C in seminal plasma, and sperm liquefaction occurred concurrently with VEGF-C activation, which was enhanced by collagen and calcium binding EGF domains 1 (CCBE1). After plasmin and ADAMTS3, KLK3 is the third protease shown to activate VEGF-C. Since differently activated VEGF-Cs are characterized by successively shorter N-terminal helices, we created an even shorter hypothetical form, which showed preferential binding to VEGFR-3. Using mass spectrometric analysis of the isolated VEGF-C-cleaving activity from human saliva, we identified cathepsin D as a protease that can activate VEGF-C as well as VEGF-D.

SUBMITTER: Jha SK

PROVIDER: S-EPMC6588350 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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KLK3/PSA and cathepsin D activate VEGF-C and VEGF-D.

Jha Sawan Kumar SK Rauniyar Khushbu K Chronowska Ewa E Mattonet Kenny K Maina Eunice Wairimu EW Koistinen Hannu H Stenman Ulf-Håkan UH Alitalo Kari K Jeltsch Michael M

eLife 20190517

Vascular endothelial growth factor-C (VEGF-C) acts primarily on endothelial cells, but also on non-vascular targets, for example in the CNS and immune system. Here we describe a novel, unique VEGF-C form in the human reproductive system produced via cleavage by kallikrein-related peptidase 3 (KLK3), aka prostate-specific antigen (PSA). KLK3 activated VEGF-C specifically and efficiently through cleavage at a novel N-terminal site. We detected VEGF-C in seminal plasma, and sperm liquefaction occur ...[more]

PMID: 31099754

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Project description:Kallikrein-related peptidase 3 (KLK3, also known as prostate-specific antigen, PSA) is a chymotrypsin-like kallikrein that has been shown to have anti-angiogenic properties. KLK3 expression in prostate cancer is associated with low microvessel density. We have previously shown in a human umbilical vein endothelial cell (HUVEC) model that the anti-angiogenic effect of KLK3 is related to its enzymatic activity. However, the mechanism of this effect remains to be clarified. To this end we have used a DNA microarray to study the KLK3-induced changes in gene expression associated with reduction of HUVEC tube formation. Among the 41000 genes studied, 311 were differentially expressed in control and KLK3-treated cells. These changes were enriched in several pathways, including the pathways associated with proteasome, ubiquitin-mediated proteolysis, focal adhesion and regulation of actin cytoskeleton. Furthermore, the changes were opposite to those described previously to occur during tubulogenesis. In conclusion, our results show that KLK3 induces gene expression changes in HUVECs. While these changes might be relevant for the mechanism by which KLK3 exerts its anti-angiogenic activity, it cannot be judged by the present results whether the changes are secondary to morphogenic differentiation of the cells or the primary mechanism mediating the effect of KLK3. Keywords: KLK3 treatment, tube formation Human umbilical vein endothelial cells (HUVECs) were isolated from umbilical cord veins and grown in Endothelial Cell Basal Medium supplemented with an Endothelial Cell Growth Medium Supplement Pack (PromoCell). The cells were cultured on Matrigel basement membrane prepara-tion together with 500 nM enzymatically active KLK3 (PSA). Phosphate buffered saline (PBS) was used for control. HUVECs were grown on Matrigel for 18 h. RNA was isolated from KLK3-treated and control HUVECs from three different individuals. Cell Recovery Solution (BD Biosciences) was used to detach cells from the Matrigel. Total RNA was isolated with the RNeasy Mini Kit (Qiagen) according to manufacturerâ??s instructions. RNA con-centration and quality of the samples was determined with an Agilent 2100 bioanalyser (Agilent Technologies) by measuring the RNA integrity number (RIN). All samples contained high quality RNA (RIN 9.5-10). DNA microarray analysis was performed with the Agilent Whole Human Genome Oligo Microar-ray 4x44K (G4112F) (Agilent Technologies Inc.) following the manufacturerâ??s protocol. Total RNA (1 ï?g) was labeled using the RNA input fluorescent linear amplification kit according to the manufacturer's recommendations (Agilent Technologies Inc.). RNA was hybridized on Agilent's Human 4x44K Oligo Microarrays containing 44000 features (41000 unique genes and transcripts). Pre-processing of the data was performed by Feature Extraction software (v 7.5.1).

Dataset Information

KLK3/PSA and cathepsin D activate VEGF-C and VEGF-D.

Publications

KLK3/PSA and cathepsin D activate VEGF-C and VEGF-D.

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