Efficient Production of Recombinant Protegrin-1 From Pichia pastoris, and Its Antimicrobial and in vitro Cell Migration Activity

Migration of capillary endothelial cells is an important component of angiogenesis in vivo. Increased numbers of mast cells have been associated with several types of angiogenesis. We have used a quantitative assay in vitro to demonstrate that mast cells release a factor that significantly increases bovine capillary endothelial cell migration. The factor is present in medium conditioned by mast cells as well as lysates of mast cells. The stimulatory effect of mast cells on migration is specific for capillary endothelial cells. Furthermore, mast cells have no mitogenic activity for capillary endothelial cells. Of all the secretory products of mast cells tested, only heparin stimulated capillary endothelial cell migration in vitro. Heparin preparations from a variety of sources stimulated capillary endothelial cell migration to the same degree but did not stimulate migration of several other cell types. The migration activity of heparin and mast cell conditioned medium was blocked by specific antagonists of heparin (protamine and heparinase), but not by chondroitinase ABC. The migration activity of mast cell conditioned medium was resistant to heat (100 degrees C) and incubation with proteolytic enzymes. These results suggest that the role of mast cells in angiogenesis may be to enhance migration of the endothelial cells of growing capillaries.

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A GDSL lipase-like from Ipomoea batatas catalyzes efficient production of 3,5-diCQA when expressed in Pichia pastoris

Communications Biology ◽

10.1038/s42003-020-01387-1 ◽

2020 ◽

Vol 3 (1) ◽

Author(s):

Sissi Miguel ◽

Guillaume Legrand ◽

Léonor Duriot ◽

Marianne Delporte ◽

Barbara Menin ◽

...

Keyword(s):

Pichia Pastoris ◽

Ipomoea Batatas ◽

Efficient Production ◽

Coffee Bean ◽

The Road ◽

Dicaffeoylquinic Acid ◽

Low Efficiency ◽

Green Coffee Bean Extract ◽

Gdsl Lipase

AbstractThe synthesis of 3,5-dicaffeoylquinic acid (3,5-DiCQA) has attracted the interest of many researchers for more than 30 years. Recently, enzymes belonging to the BAHD acyltransferase family were shown to mediate its synthesis, albeit with notably low efficiency. In this study, a new enzyme belonging to the GDSL lipase-like family was identified and proven to be able to transform chlorogenic acid (5-O-caffeoylquinic acid, 5-CQA, CGA) in 3,5-DiCQA with a conversion rate of more than 60%. The enzyme has been produced in different expression systems but has only been shown to be active when transiently synthesized in Nicotiana benthamiana or stably expressed in Pichia pastoris. The synthesis of the molecule could be performed in vitro but also by a bioconversion approach beginning from pure 5-CQA or from green coffee bean extract, thereby paving the road for producing it on an industrial scale.

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Malotilate reduces collagen synthesis and cell migration activity of fibroblasts in vitro

Biochemical Pharmacology ◽

10.1016/0006-2952(87)90464-3 ◽

1987 ◽

Vol 36 (22) ◽

pp. 3957-3963 ◽

Cited By ~ 1

Author(s):

A. Poeschl ◽

D. Rehn ◽

J.-M. Dumont ◽

P.K. Mueller ◽

G. Hennings

Keyword(s):

Cell Migration ◽

Collagen Synthesis ◽

Migration Activity

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THE EFFECT OF GLUTAMATE ON THE MIGRATION OF T CELLS FROM HEALTHY DONORS AND PATIENTS WITH MULTIPLE SCLEROSIS IN VITRO

Medical academic journal ◽

10.17816/maj191s129-30 ◽

2019 ◽

Vol 19 (1S) ◽

pp. 29-30

Author(s):

M A Maksimova ◽

U Sh Kuzmina ◽

K Z Bakhtiyarova ◽

Yu V Vakhitova

Keyword(s):

Multiple Sclerosis ◽

T Cells ◽

Cell Migration ◽

T Cell ◽

Concentration Gradient ◽

Migration Activity ◽

T Cell Migration ◽

Healthy Donors ◽

Glutamate Receptor Agonists

Aim of study. To study chemotactic properties of glutamate and glutamate receptor agonists on T cells migration from healthy donors and patients with multiple sclerosis (MS) in vitro. Materials and methods. T cell migration of 15 patients with MS and 15 healthy donors was studied in vitro using transwells. Lymphocytes were activated with PMA (10 ng/mL). T cells were added to transwells with fibronectin (10 μg/mL) pretreated membrane. The lower chamber contained glutamate or AMPA or NMDA (100 μM for each) in complete RPMI medium. Migrated cells were collected and stained with antibodies to CD3-marker for subsequent analysis by cytofluorimetry. Results and conclusion. In presence of glutamate, there is a tendency to a decrease in migration activity in both groups of donors. T-cell chemotaxis of healthy donors, but not MS patients, decreased in concentration gradient of NMDA. The activation of lymphocytes with PMA leads to a decrease in the number of migrated cells by an average of 17% (p < 0.01). In MS patients there is a tendency to an increase in chemotaxis of activated cells in concentration gradient of glutamate, and a decrease with AMPA. Thus, glutamate and glutamate receptors agonists do not possess pronounced chemotactic properties, but rather enhance T-cell migration through synthesis of adhesion molecules on the surface of lymphocytes and endothelium.

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Neutrophil migration through in vitro interstitial matrix

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100124872 ◽

1992 ◽

Vol 50 (1) ◽

pp. 894-895

Author(s):

J. Roemer ◽

S.R. Simon

Keyword(s):

Extracellular Matrix ◽

Smooth Muscle ◽

Cell Migration ◽

Smooth Muscle Cells ◽

Inflammatory Cell ◽

Neutrophil Migration ◽

Culture Conditions ◽

Aortic Smooth Muscle ◽

Specific Culture

We are developing an in vitro interstitial extracellular matrix (ECM) system for study of inflammatory cell migration. Falcon brand Cyclopore membrane inserts of various pore sizes are used as a support substrate for production of ECM by R22 rat aortic smooth muscle cells. Under specific culture conditions these cells produce a highly insoluble matrix consisting of typical interstitial ECM components, i.e.: types I and III collagen, elastin, proteoglycans and fibronectin.

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The Migration of Human Smooth Muscle Cells In Vitro Is Mediated by Plasminogen Activation and Can Be Inhibited by α2-Macroglobulin Receptor Associated Protein

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657646 ◽

1997 ◽

Vol 78 (02) ◽

pp. 880-886 ◽

Cited By ~ 46

Author(s):

Monique J Wijnberg ◽

Paul H A Quax ◽

Nancy M E Nieuwenbroek ◽

Jan H Verheijen

Keyword(s):

Smooth Muscle ◽

Cell Migration ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Ldl Receptor ◽

Migration And Invasion ◽

Invasion Assay ◽

Plasminogen Activation ◽

Smooth Muscle Cell Migration

SummaryThe plasminogen activation system is thought to be important in cell migration processes. A role for this system during smooth muscle cell migration after vascular injury has been suggested from several animal studies. However, not much is known about its involvement in human vascular remodelling. We studied the involvement of the plasminogen activation system in human smooth muscle cell migration in more detail using an in vitro wound assay and a matrix invasion assay. Inhibition of plasmin activity or inhibition of urokinase-type plasminogen activator (u-PA) activity resulted in approximately 40% reduction of migration after 24 h in the wound assay and an even stronger reduction (70-80%) in the matrix invasion assay. Migration of smooth muscle cells in the presence of inhibitory antibodies against tissue-type plasminogen activator (t-PA) was not significantly reduced after 24 h, but after 48 h a 30% reduction of migration was observed, whereas in the matrix invasion assay a 50% reduction in invasion was observed already after 24 h. Prevention of the interaction of u-PA with cell surface receptors by addition of soluble u-PA receptor or α2-macroglobulin receptor associated protein (RAP) to the culture medium, resulted in a similar inhibition of migration and invasion. From these results it can be concluded that both u-PA and t-PA mediated plasminogen activation can contribute to in vitro human smooth muscle cell migration and invasion. Furthermore, the interaction between u-PA and its cell surface receptor appears also to be involved in this migration and invasion process. The inhibitory effects on migration and invasion by the addition of RAP suggests an involvement of a RAP sensitive receptor of the LDL receptor family, possibly the LDL-receptor related protein (LRP) and/or the VLDL receptor.

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CXCR4-targeted Nanoparticles Reduce Cell Viability, Induce Apoptosis and Inhibit SDF-1α Induced BT-549-Luc Cell Migration In Vitro

Current Drug Delivery ◽

10.2174/1567201814666170216130448 ◽

2017 ◽

Vol 14 (8) ◽

Cited By ~ 1

Author(s):

Chuda Chittasupho ◽

Prartana Kewsuwan ◽

Takashi Murakami

Keyword(s):

Cell Migration ◽

Cell Viability ◽

Targeted Nanoparticles ◽

Reduce Cell Viability

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In vitro Tumor Cell Migration Assay Using ThinCertsTM (Transwells)

BIO-PROTOCOL ◽

10.21769/bioprotoc.1830 ◽

2016 ◽

Vol 6 (11) ◽

Cited By ~ 1

Author(s):

Marc Schneider

Keyword(s):

Cell Migration ◽

Tumor Cell ◽

Cell Migration Assay ◽

Migration Assay ◽

Tumor Cell Migration

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Pals1 prevents Rac1-dependent colorectal cancer cell metastasis by inhibiting Arf6

Molecular Cancer ◽

10.1186/s12943-021-01354-2 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Simona Mareike Lüttgenau ◽

Christin Emming ◽

Thomas Wagner ◽

Julia Harms ◽

Justine Guske ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Migration ◽

Cancer Cells ◽

Cancer Cell ◽

Scaffolding Protein ◽

Migration And Invasion ◽

Tight Junction Formation ◽

Cell Metastasis ◽

Colorectal Cancer Cells

AbstractLoss of apical-basal polarity and downregulation of cell-cell contacts is a critical step during the pathogenesis of cancer. Both processes are regulated by the scaffolding protein Pals1, however, it is unclear whether the expression of Pals1 is affected in cancer cells and whether Pals1 is implicated in the pathogenesis of the disease.Using mRNA expression data and immunostainings of cancer specimen, we show that Pals1 is frequently downregulated in colorectal cancer, correlating with poorer survival of patients. We further found that Pals1 prevents cancer cell metastasis by controlling Rac1-dependent cell migration through inhibition of Arf6, which is independent of the canonical binding partners of Pals1. Loss of Pals1 in colorectal cancer cells results in increased Arf6 and Rac1 activity, enhanced cell migration and invasion in vitro and increased metastasis of transplanted tumor cells in mice. Thus, our data reveal a new function of Pals1 as a key inhibitor of cell migration and metastasis of colorectal cancer cells. Notably, this new function is independent of the known role of Pals1 in tight junction formation and apical-basal polarity.

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