Deoxyhypusine synthase catalyzes the first step in posttranslational synthesis of an unusual amino acid, hypusine (N epsilon-(4-amino-2-hydroxybutyl)lysine), eukaryotic translation initiation factor 5A (eIF-5A) precursor protein. The null mutation single copy gene, yDHS, encoding deoxyhypusine results loss viability yeast Saccharomyces cerevisiae. Upon depletion synthase, and consequently eIF-5A, cessation growth was accompanied by a marked enlargement cells, suggesting defect cell cycle...

Abstract Mesenchymal stromal/stem cells (MSCs) have been developed as a promising source for cell-based therapies of ischemic disease. However, there are some hurdles in their clinical application such poor cell engraftment and inconsistent stem potency. In this study, we sought to find biomarkers predicting potency MSCs proangiogenic therapy improve beneficial effects. Large variations were observed factor secretion profiles conditioned media derived from nine different donor-derived...

Silicon dioxide composites have been found to enhance the mechanical properties of scaffolds and support growth human adipose tissue-derived stem cells (hADSCs) both in vitro vivo. Silica (silicon alone) exists as differently sized particles when suspended culture medium, but it is not clear whether particle size influences beneficial effect silicon on hADSCs. In this study, we examined different mitogen-activated protein kinase signaling hADSCs.Silica gel was prepared by a chemical reaction...

Deoxyhypusine synthase catalyzes the first step in post-translational formation of hypusine (N epsilon-(4-amino-2-hydroxybutyl)lysine). cDNA clones encoding deoxyhypusine were isolated from a human HeLa cell library. Full-length 369-amino acid protein (calculated molecular mass 40,970 Da) and shorter clone that would potentially encode with an internal deletion 56 amino acids (Asp262-Ser317) isolated. The deduced sequence enzyme shows high degree identity to yeast known sequences tryptic...

Peroxidasin (PXDN) has been reported to crosslink the C-terminal non-collagenous domains of collagen IV (Col IV) by forming covalent sulfilimine bond. Here, we explored physiological role PXDN and its mechanism action in endothelial cell survival growth. Silencing using siRNAs decreased proliferation without increase number detached cells viability under serum-starved condition with increased fragmented nuclei caspase 3/7 activity. Conditioned medium (CM) containing wild-type restored...

Urokinase plasminogen activator (uPA) belongs to a family of proteins that contains kringle domain and plays an important role in inflammation, tissue remodeling, angiogenesis, tumor metastasis by pericellular activation. Kringle domains have been shown demonstrate anti-angiogenic anti-tumor activities. Here, we report our investigation the uPA for activity possible cellular mechanism action. The recombinant (Asp⁴⁵-Lys¹³⁵) (UK1) inhibited endothelial cell proliferation...

The alkylating agent temozolomide (TMZ) is an effective drug used for the treatment of malignant gliomas. However, tumor relapse combined with development resistance remains a significant problem. To clarify mechanism glioma cells to TMZ chemotherapy, TMZ-resistant cell lines (TR cells) were generated using U373 and U251 human cells, TMZ-resistance was confirmed via viability apoptosis assays. TR not associated molecule O(6)-methylguanine-DNA-methyltransferase. Notably, expression level...

Endothelial progenitor cells (EPCs) have been shown to therapeutic potential in ischemic disease. However, the number of EPCs for cell therapy is limited. In this study, instead typical adherent culture method, we investigated a more efficient, clinically applicable nonadhesive expansion method early using cord blood-derived overcome rapid cellular senescence. After suspension isolated CD34(+) serum-free medium containing each cytokine combination was maintained 9 d, expanded functional...

Mebendazole (MBZ), a microtubule depolymerizing drug commonly used for the treatment of helminthic infections, has recently been noted as repositioning candidate angiogenesis inhibition and cancer therapy. However, definite anti-angiogenic mechanism MBZ remains unclear. In this study, we explored inhibitory in endothelial cells (ECs) developed novel strategy to improve its Treatment ECs with led EC proliferation dose-dependent manner several culture conditions presence vascular growth factor...

Deoxyhypusine synthase catalyses the first step in post-translational synthesis of hypusine [Nepsilon-(4-amino-2-hydroxybutyl) lysine] a single cellular protein, precursor eukaryotic initiation factor 5A (eIF5A). exists as tetramer with four potential active sites. The formation stable complex between human deoxyhypusine and its protein substrate, recombinant eIF5A (ec-eIF5A), was examined by affinity chromatography using polyhistidine-tagged (His.Tag) ec-eIF5A, gel mobility-shift method,...

Mesenchymal stromal/stem cells (MSCs) have the potential to differentiate into neuron-like under specific conditions and secrete paracrine factors for neuroprotection regeneration. Previously, Rho-kinase inhibitors been reported potentiate differentiation of rodent bone marrow MSCs induced by CoCl2 (HIF-1α activation-mimicking agent). Here, a strategy priming with fasudil, inhibitor, was investigated using Wharton's jelly-derived (WJ-MSCs) improve recovery in rat model intracranial...

The serine protease urokinase-type plasminogen activator (uPA) is implicated in pericellular proteolysis a variety of physiological and pathological processes including angiogenesis tumor metastasis. kringle domain uPA (UK1) has proven to be an anti-angiogenic molecule with unknown mechanism amino terminal fragment (u-ATF) additional growth factor-like can used for blocking interaction receptor. Here, we compared activities these two molecules vitro vivo. recombinant u-ATF from E. coli...

Neovascularization plays a critical role in the growth and metastatic spread of tumors involves recruitment circulating endothelial progenitor cells (EPC) from bone marrow as well sprouting preexisting cells. In this study, we examined if EPCs could promote tumor angiogenesis would be an effective cellular target for inhibitor, recombinant kringle domain tissue-type plasminogen activator (TK1-2). When TK1-2 was applied ex vivo culture isolated human cord blood, inhibited adhesive...

Peroxidasin (PXDN) is a unique peroxidase containing extracellular matrix motifs and stabilizes collagen IV networks by forming sulfilimine crosslinks. PXDN gene knockout in Caenorhabditis elegans (C. elegans) Drosophila results the demise at embryonic larval stages. mutations lead to severe eye disorders, including microphthalmia, cataract, glaucoma, anterior segment dysgenesis humans mice. To investigate how loss of function affects organ development, we generated Pxdn mice deletion exon 1...

Abstract The recombinant two kringle domain of human tissue-type plasminogen activator (TK1-2) has been shown to inhibit endothelial cell proliferation, angiogenesis, and tumor growth despite sharing a low amino acid sequence homology with angiostatin. Here, we explored possible inhibitory mechanism action TK1-2 by focusing on antimigratory effect. effectively inhibited migration induced basic fibroblast factor or vascular in dose-dependent manner tube formation Matrigel. It blocked...

Deoxyhypusine synthase catalyzes the first step in post-translational synthesis of hypusine [Nε-(4-amino-2-hydroxybutyl)lysine] eukaryotic translation initiation factor 5A. We recently reported biochemical evidence for a covalent enzyme-substrate intermediate involving specific lysine residue (Lys329) human deoxyhypusine (Wolff, E. C., Folk, J. E., and Park, M. H. (1997) Biol. Chem. 272, 15865–15871). In an effort to evaluate role this catalysis, we carried out site-directed mutagenesis (Lys...