scholarly journals Recombinant Sip Young 6 His Protein Production from the Sea Star Igkappa Gene, after Cloning

2021 ◽  
Vol 2 (2) ◽  
pp. 85-86
Author(s):  
Michel Leclerc
Keyword(s):  
Protein Production ◽  
Sea Star ◽  
Hek 293 ◽  
First Time

In 2014 we have isolated and cloned the sea star Igkappa gene which induced an anti-HRP primitive invertebrate antibody. We attempt, in the present work, for the first time in Invertebrates, to produce a SIP Young 6 His protein through HeK 293 EBNA cells.

scholarly journals Phosphoregulation of the intracellular termini of K+-Cl− cotransporter 2 (KCC2) enables flexible control of its activity

2018 ◽  
Vol 293 (44) ◽  
pp. 16984-16993 ◽  
Author(s):  
Antje Cordshagen ◽  
Wiebke Busch ◽  
Michael Winklhofer ◽  
Hans Gerd Nothwang ◽  
Anna-Maria Hartmann
Keyword(s):  
Intrinsic Activity ◽  
Hek 293 ◽  
Flexible Control ◽  
Mediated Effects ◽  
293 Cells ◽  
Bona Fide ◽  
Graded Activity ◽  
First Time ◽  
Increased Activity

The pivotal role of K+-Cl− cotransporter 2 (KCC2) in inhibitory neurotransmission and severe human diseases fosters interest in understanding posttranslational regulatory mechanisms such as (de)phosphorylation. Here, the regulatory role of the five bona fide phosphosites Ser31, Thr34, Ser932, Thr999, and Thr1008 was investigated by the use of alanine and aspartate mutants. Tl+-based flux analyses in HEK-293 cells demonstrated increased transport activity for S932D (mimicking phosphorylation) and T1008A (mimicking dephosphorylation), albeit to a different extent. Increased activity was due to changes in intrinsic activity, as it was not caused by increased cell-surface abundance. Substitutions of Ser31, Thr34, or Thr999 had no effect. Additionally, we show that the indirect actions of the known KCC2 activators staurosporine and N-ethylmaleimide (NEM) involved multiple phosphosites. S31D, T34A, S932A/D, T999A, or T1008A/D abrogated staurosporine mediated stimulation, and S31A, T34D, or S932D abolished NEM-mediated stimulation. This demonstrates for the first time differential effects of staurosporine and NEM on KCC2. In addition, the staurosporine-mediated effects involved both KCC2 phosphorylation and dephosphorylation with Ser932 and Thr1008 being bona fide target sites. In summary, our data reveal a complex phosphoregulation of KCC2 that provides the transporter with a toolbox for graded activity and integration of different signaling pathways.

scholarly journals Cecropia pachystachya Leaves Present Potential to Be Used as New Ingredient for Antiaging Dermocosmetics

2019 ◽  
Vol 2019 ◽  
pp. 1-9
Author(s):  
Maria Fernanda Fernandes ◽  
Jessica Leiras Mota Conegundes ◽  
Nícolas de Castro Campos Pinto ◽  
Luiz Gustavo de Oliveira ◽  
Jair Adriano Kopke de Aguiar ◽  
...  
Keyword(s):  
Biological Activities ◽  
Thiobarbituric Acid ◽  
Skin Aging ◽  
Total Phenolic ◽  
Leaf Extracts ◽  
Hek 293 ◽  
Human Embryonic Kidney Cells ◽  
Glycation End Products ◽  
First Time

Several biological activities have been reported for leaf extracts of Cecropia pachystachya species, including antioxidant and wound healing activities. This study aims to report, for the first time, the antiaging potential of the hydroethanolic (HE) and the ethanolic (EE) extracts obtained from the leaves of C. pachystachya using different in vitro assays. Both HE and EE presented relevant antioxidant capacity in different models, including phosphomolybdenum, 1,1-diphenyl-2-picryl-hydrazyl (DPPH), carotene/linoleic acid bleaching, and thiobarbituric acid reactive substances (TBARS) assays. Their ability to prevent the production of advanced glycation end products (AGEs) was also evaluated, and both extracts showed important activity, especially HE. The extracts also stimulated the fibroblasts proliferation in vitro, specialized cells that produce several mediators which maintain the skin integrity and youthfulness. Cytotoxicity of the extracts was not observed for this lineage or HEK-293, human embryonic kidney cells widely used to evaluate cytotoxicity of chemical compounds. HE also exhibited the ability to inhibit the collagenase (metalloproteinase MMP-2) and elastase activities. The total phenolic and flavonoids contents were also determined. HPLC analysis revealed the presence of the flavonoids orientin and iso-orientin, which were quantified to be used as chemical markers. The results suggested that the extracts of C. pachystachya leaves present the potential to be used in dermocosmetic formulations to prevent the skin aging process, which attracts the attention of pharmaceutical companies and researchers interested in the development of novel ingredients likely to be used as active principles in antiaging products.

scholarly journals Identifying HIPK1 as Target of miR-22-3p Enhancing Recombinant Protein Production From HEK 293 Cell by Using Microarray and HTP siRNA Screen

2017 ◽  
Vol 13 (2) ◽  
pp. 1700342 ◽  
Author(s):  
Sarah Inwood ◽  
Eugen Buehler ◽  
Michael Betenbaugh ◽  
Madhu Lal ◽  
Joseph Shiloach
Keyword(s):  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Sirna Screen ◽  
Hek 293

Determination of apparent calcium affinity for endogenously expressed human sarco(endo)plasmic reticulum calcium-ATPase isoform SERCA3

2009 ◽  
Vol 296 (5) ◽  
pp. C1105-C1114 ◽  
Author(s):  
P. Charukeshi Chandrasekera ◽  
Margaret E. Kargacin ◽  
Julie P. Deans ◽  
Jonathan Lytton
Keyword(s):  
Cell Types ◽  
Protein Isoforms ◽  
Inhibitory Antibody ◽  
Human Tonsil ◽  
Hek 293 ◽  
293 Cells ◽  
Plasmic Reticulum ◽  
Two Component ◽  
Calcium Affinity ◽  
First Time

The sarco(endo)plasmic reticulum Ca2+-ATPases (SERCAs) play a crucial role in regulating free cytosolic Ca2+ concentration in diverse cell types. It has been shown that recombinant SERCA3, when measured in heterologous systems, exhibits low apparent affinity for Ca2+; however, Ca2+ affinity of native SERCA3 in an endogenous setting has not been examined. Such a measurement is complicated, because SERCA3 is always coexpressed with the housekeeping isoform SERCA2b. We used a fluorescence-based assay for monitoring continuous Ca2+ uptake into microsomes to examine the properties of endogenous human SERCA3 and SERCA2b. The kinetic parameters were derived using a cooperative two-component uptake model for Ca2+ activation, and the values assigned to SERCA3 were confirmed using the highly specific human SERCA3 inhibitory antibody PL/IM430. First, using recombinant human SERCA3 and SERCA2b proteins transiently expressed in HEK-293 cells, we confirmed the previously observed low apparent Ca2+ affinity for SERCA3 compared with SERCA2b (1.10 ± 0.04 vs. 0.26 ± 0.01 μM), and using mixtures of recombinant protein isoforms, we validated the two-component uptake model. Then we determined apparent Ca2+ affinity for SERCA proteins present endogenously in cultured Jurkat T lymphocytes and freshly isolated human tonsil lymphocytes. The apparent Ca2+ affinity in these two preparations was 1.04 ± 0.07 and 1.1 ± 0.2 μM for SERCA3 and 0.27 ± 0.02 and 0.26 ± 0.01 μM for SERCA2b, respectively. Our data demonstrate, for the first time, that affinity for Ca2+ is inherently lower for SERCA3 expressed in situ than for other SERCA isoforms.

scholarly journals Influence of glutamine on transient and stable recombinant protein production in CHO and HEK-293 cells

2011 ◽  
Vol 5 (Suppl 8) ◽  
pp. P35 ◽  
Author(s):  
Yashas Rajendra ◽  
Divor Kiseljak ◽  
Lucia Baldi ◽  
David L Hacker ◽  
Florian M Wurm
Keyword(s):  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

scholarly journals New Mediterranean Biodiversity Records (December 2017)

2018 ◽  
Vol 18 (3) ◽  
pp. 534 ◽  
Author(s):  
COLLECTIVE ARTICLE A
Keyword(s):  
New Record ◽  
New Records ◽  
Marine Species ◽  
First Record ◽  
Tyrrhenian Sea ◽  
Sea Star ◽  
First Case ◽  
The Mediterranean ◽  
First Time ◽  
Mediterranean Biodiversity

The “New Mediterranean Biodiversity Records” series includes new records of marine species found in the Mediterranean basin and/or information on the spatial distribution of already established species of particular interest. The current article presents information on 20 marine taxa classified per country according to their geographic position in the Mediterranean, from west to east. The new records per ecoregion are as follows: Algeria: the first record of the fish Etrumeus golanii is reported along the Algerian coast. Tunisia: the alien jellyfish Phyllorhiza punctata is reported for the first time in the Gulf of Gabès. Italy: the first record of Siganus rivulatus in the Strait of Sicily and a new record of Katsuwonus pelamis from the central Tyrrhenian Sea are reported. The establishment of the isopod of the genus Mesanthura in the northern Tyrrhenian with some notes on its ecology are also included. Croatia: signs of establishment of the Lessepsian Siganus luridus and the occurrence of the alien mollusc Rapana venosa are reported. Albania: the first record of the elasmobranch Alopias superciliosus and a recent sighting of the rare monk seal Monachus monachus in Albanian waters are given. Greece: signs of the establishment of the fish Parupeneus forsskali and of the ascidian Hermania momus in Hellenic Aegean waters are reported. Turkey: a new record of the fish P. forsskali and of the Acarea of the genus Acaromantis and Simognathus are given, while the first case of Remora australis in association with delphinids and the occurrence of the sea star Coscinasterias tenuispina are reported. Also, the establishment of the two alien species Isognomon legumen and Viriola sp. [cf. corrugata] are presented. Egypt: the fish Bathygobius cyclopterus is reported for the first time in Mediterranean Sea waters. Also, a new record of Pagellus bogaraveo and a first record of Seriola fasciata in Egyptian Mediterranean waters are reported. Lebanon: the first record of Dondice banyulensis is presented.

Annexin 1 and its bioactive peptide inhibit neutrophil-endothelium interactions under flow: indication of distinct receptor involvement

Blood ◽  
2006 ◽  
Vol 107 (5) ◽  
pp. 2123-2130 ◽  
Author(s):  
Richard P. G. Hayhoe ◽  
Ahmad M. Kamal ◽  
Egle Solito ◽  
Roderick J. Flower ◽  
Dianne Cooper ◽  
...  
Keyword(s):  
Flow Chamber ◽  
Mechanisms Of Action ◽  
Hek 293 ◽  
Erk Activation ◽  
Transfected Cells ◽  
Annexin 1 ◽  
293 Cells ◽  
Neutralizing Monoclonal Antibody ◽  
Human Pmns ◽  
First Time

We have tested the effects of annexin 1 (ANXA1) and its N-terminal peptide Ac2-26 on polymorphonuclear leukocyte (PMN) recruitment under flow. Differential effects of the full-length protein and its peptide were observed; ANXA1 inhibited firm adhesion of human PMNs, while Ac2-26 significantly attenuated capture and rolling without effect on firm adhesion. Analysis of the effects of ANXA1 and Ac2-26 on PMN adhesion molecule expression supported the flow chamber results, with Ac2-26 but not ANXA1 causing l-selectin and PSGL-1 shedding. ANXA1 and its peptide act via the FPR family of receptors. This was corroborated using HEK-293 cells transfected with FPR or FPRL-1/ALX (the 2 members of this family expressed by human PMNs). While Ac2-26 bound both FPR and FPRL-1/ALX, ANXA1 bound FPRL-1/ALX only. ANXA1 and Ac2-26 acted as genuine agonists; Ac2-26 binding led to ERK activation in both FPR- and FPRL-1/ALX-transfected cells, while ANXA1 caused ERK activation only in cells transfected with FPRL-1/ALX. Finally, blockade of FPRL-1/ALX with a neutralizing monoclonal antibody was found to abrogate the effects of ANXA1 in the flow chamber but was without effect on Ac2-26-mediated inhibition of rolling. These findings demonstrate for the first time distinct mechanisms of action for ANXA1 and its N-terminal peptide Ac2-26.

scholarly journals The Sea Star Igkappa Gene and The Ophuirid Igkappa Gene Vaccinations to HRP (Horse-radish Peroxydase)

2019 ◽  
Vol 4 (3) ◽  
Keyword(s):  
Complement System ◽  
Receptor Gene ◽  
Fc Receptor ◽  
Sea Star ◽  
Asterias Rubens ◽  
Genomic Studies ◽  
First Time ◽  
The Complement System

The main point of the sea star immunology, of the ophuirid immunology remain the discovery of the INVERTEBRATE PRIMITIVE ANTIBODY (IPA), the IGKAPPA GENES, with Ig sites which imply the COMPLEMENT SYSTEM, to be initiated: 9 component genes from C1 to C9 have been updated these last years, in sea star genome, in ophuirid one. These primitive antibodies were obtained after immunizations to the enzyme HRP (Horse-radish peroxydase). It is the first time we observe vaccinations in invertebrates. We have discovered, for the first time also, in these same Invertebrate a FC receptor gene, a Fab gene which corroborate the presence of IPA. The transcriptomes are given. It is the first time, we can speak of Adaptative Immunity, in Echinoderms, in Invertebrates Since many years, even since a century long, the notion of Antibody was out of the speech of immunologists. To speak of that made you as an outlaw. It is time to look with genomic studies which confirm which assert now evidence that 3 classes of Echinoderms out of 5 possess an Igkappa gene, a FC receptor gene. I recall these classent: the Asterids with Asterias rubens, the Ophuirids with Ophiocomina nigra and the Crinoïds.

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