keyword
MENU ▼
Read by QxMD icon Read
search

enterokinase

keyword
https://www.readbyqxmd.com/read/29629340/enterokinase-enhances-influenza-a-virus-infection-by-activating-trypsinogen-in-human-cell-lines
#1
Hideki Hayashi, Yoshinao Kubo, Mai Izumida, Etsuhisa Takahashi, Hiroshi Kido, Ko Sato, Mutsuo Yamaya, Hidekazu Nishimura, Kou Nakayama, Toshifumi Matsuyama
Cleavage and activation of hemagglutinin (HA) by trypsin-like proteases in influenza A virus (IAV) are essential prerequisites for its successful infection and spread. In host cells, some transmembrane serine proteases such as TMPRSS2, TMPRSS4 and HAT, along with plasmin in the bloodstream, have been reported to cleave the HA precursor (HA0 ) molecule into its active forms, HA1 and HA2 . Some trypsinogens can also enhance IAV proliferation in some cell types (e.g., rat cardiomyoblasts). However, the precise activation mechanism for this process is unclear, because the expression level of the physiological activator of the trypsinogens, the TMPRSS15 enterokinase, is expected to be very low in such cells, with the exception of duodenal cells...
2018: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/29506820/tandem-repeated-expression-of-lunasin-gene-in-pichia-pastoris-and-its-anti-inflammatory-activity-in-vitro
#2
Yingying Zhu, Everaert Nadia, Yang Yao, Zhenxing Shi, Guixing Ren
Lunasin is a novel promising health-beneficial peptide derived from soybean. However, the application of lunasin is limited by its high cost. In this study, we developed a successful protocol for expression of a dimer formation protein containing 4 tandem repeated lunasin analogs (lunasin-4) in Pichia pastoris. The expression level at the optimal condition (initial pH 7.0, 1.0% final methanol concentration and induction for 72 h at 26 °C) was 0.24 mg/mL cell-free broth. Lunasin analog, obtained from purified lunasin-4 protein through enterokinase digestion and ultrafiltration, significantly decreased (p < 0...
March 2, 2018: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/29408294/efficient-expression-and-isolation-of-recombinant-human-interleukin-11-rhil-11-in-pichia-pastoris
#3
Kuo-Ming Yu, Johnson Yiu-Nam Lau, Manson Fok, Yuk-Keung Yeung, Siu-Ping Fok, Felix Shek, Wing-Tak Wong, Qui-Lim Choo
Current source of recombinant human interleukin-11 (rhIL-11) is isolated from a fusion protein expressed by E. coli that requires additional enterokinase to remove linked protein, resulting in product heterogeneity of N-terminal sequence. Due to lack of glycosylation, rhIL-11 is suitable to be expressed by yeast cells. However, the only available yeast-derived rhIL-11 presents an obstacle in low production yield, as well as an unamiable process, such as the use of reverse-phase chromatography employing plenty of toxic organic solvents...
February 2, 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/29369519/molecular-farming-in-barley-development-of-a-novel-production-platform-to-produce-human-antimicrobial-peptide-ll-37
#4
Edita Holásková, Petr Galuszka, Alžbeta Mičúchová, Marek Šebela, M Tufan Öz, Ivo Frébort
The peptide LL-37, a component of the human innate immune system, represents a promising drug candidate. In particular, the development of low-cost production platform technology is a critical bottleneck in its use in medicine. In the present study, a viable approach for the LL-37 production in transgenic barley was developed. First, comparative analyses of the effects of different fused peptide epitope tags applicable for accumulation and purification on LL-37 production yield were performed using transient expression in tobacco leaves...
January 25, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29349619/-1-h-15-n-and-13-c-chemical-shift-assignments-of-the-micelle-immersed-fat-c-terminal-fatc-domains-of-the-human-protein-kinases-ataxia-telangiectasia-mutated-atm-and-dna-dependent-protein-kinase-catalytic-subunit-dna-pkcs-fused-to-the-b1-domain-of-streptococcal
#5
Munirah S Abd Rahim, Lisa A M Sommer, Anja Wacker, Martin Schaad, Sonja A Dames
FAT C-terminal (FATC) is a circa 33 residue-long domain. It controls the kinase functionality in phosphatidylinositol-3 kinase-related kinases (PIKKs). Recent NMR- and CD-monitored interaction studies indicated that the FATC domains of all PIKKs can interact with membrane mimetics albeit with different preferences for membrane properties such as surface charge and curvature. Thus they may generally act as membrane anchoring unit. Here, we present the 1 H, 15 N, and 13 C chemical shift assignments of the DPC micelle immersed FATC domains of the human PIKKs ataxia-telangiectasia mutated (ATM, residues 3024-3056) and DNA protein kinase catalytic subunit (DNA-PKcs, residues 4096-4128), both fused to the 56 residue long B1 domain of Streptococcal protein G (GB1)...
April 2018: Biomolecular NMR Assignments
https://www.readbyqxmd.com/read/29328651/enzymatic-cleavage-of-branched-peptides-for-targeting-mitochondria
#6
Hongjian He, Jiaqing Wang, Huaimin Wang, Ning Zhou, Dongsik Yang, Douglas R Green, Bing Xu
Most of the reported mitochondria-targeting molecules are lipophilic and cationic, and thus they may become cytotoxic with accumulation. Here we show enzymatic cleavage of branched peptides that carry negative charges for targeting mitochondria. Conjugating a well-established protein tag (i.e., FLAG-tag) to self-assembling motifs affords the precursors that form micelles. Enzymatic cleavage of the hydrophilic FLAG motif (DDDDK) by enterokinase (ENTK) turns the micelles to nanofibers. After being taken up by cells, the micelles, upon the action of intracellular ENTK, turn into nanofibers to locate mainly at mitochondria...
January 31, 2018: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29223155/recombinant-human-erythropoietin-with-additional-processable-protein-domains-purification-of-protein-synthesized-in-escherichia-coli-heterologous-expression-system
#7
T M Grunina, A V Demidenko, A M Lyaschuk, M S Poponova, Z M Galushkina, L A Soboleva, S A Cherepushkin, N B Polyakov, D A Grumov, A I Solovyev, V G Zhukhovitsky, I S Boksha, M E Subbotina, A V Gromov, V G Lunin, A S Karyagina
Three variants of human recombinant erythropoietin (rhEPO) with additional N-terminal protein domains were obtained by synthesis in an Escherichia coli heterologous expression system. These domains included (i) maltose-binding protein (MBP), (ii) MBP with six histidine residues (6His) in N-terminal position, (iii) s-tag (15-a.a. oligopeptide derived from bovine pancreatic ribonuclease A) with N-terminal 6His. Both variants of the chimeric protein containing MBP domain were prone to aggregation under nondenaturing conditions, and further purification of EPO after the domain cleavage by enterokinase proved to be impossible...
November 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/29037914/antimicrobial-activity-and-preliminary-mode-of-action-of-plnef-expressed-in-escherichia-coli-against-staphylococci
#8
Xuan Tang, Shuangshuang Wu, Xiaofeng Wang, Qing Gu, Ping Li
Strains belonging to the genus of Staphylococci, such as Staphylococcus aureus are common pathologic bacteria which may cause nosocomial cross infection and food contamination. Plantaricin EF (PlnEF), a two-peptide nonlantibiotic bacteriocin produced by Lactobacillus plantarum strains, shows great inhibitory effects against Gram-positive Staphylococci strains. To overproduce this two-peptide bacteriocin, plnE and plnF genes were cloned into pET32a (+) vector and heterologously expressed in Escherichia coli by fusion with His6-tag in this study...
March 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/28988146/recombinant-production-of-the-insecticidal-scorpion-toxin-bj%C3%AE-it-in-escherichia-coli
#9
Hongbo Li, Yuxian Xia
Scorpion long-chain insect neurotoxins have important potential application value in agricultural pest control. The difficulty of obtaining natural toxins is the major obstacle preventing analyses of their insecticidal activity against more agricultural insect pests. Here we cloned the insect neurotoxin BjαIT gene into the pET32 expression vector and expressed the resulting thioredoxin (Trx)-BjαIT fusion protein in Escherichia coli. Soluble Trx-BjαIT was expressed at a high level when induced at 18 °C with 0...
February 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/28902573/preparation-of-umami-octopeptide-with-recombined-escherichia-coli-feasibility-and-challenges
#10
Liming Zhao, Yin Zhang, Chandrasekar Venkitasamy, Zhongli Pan, Longyi Zhang, Siya Guo, Wei Xiong, Hu Xia, Liu Wenlong, Gou Xinhua
The taste of umami peptide H-Lys-Gly-Asp-Glu-Glu-Ser-Leu-Ala-OH (LGAGGSLA) is controversial. One possible reason for this controversy is the use of chemically synthesized LGAGGSLA to confirm its taste. To explore other ways to further confirm the flavor of LGAGGSLA, we developed a new strategy to prepare a bio-source peptide by adopting a gene engineering method to express LGAGGSLA in recombinant Escherichia coli. In our previous work, we structured the LGAGGSLA recombinant expression system and optimized the culturing conditions for preparing a fusion protein...
September 13, 2017: Bioengineered
https://www.readbyqxmd.com/read/28734839/recombinant-expression-of-ixolaris-a-kunitz-type-inhibitor-from-the-tick-salivary-gland-for-nmr-studies
#11
V S De Paula, F H S Silva, I M B Francischetti, R Q Monteiro, A P Valente
Ixolaris is an anticoagulant protein identified in the tick saliva of Ixodes scapularis. Ixolaris contains 2 Kunitz like domains and binds to Factor Xa or Factor X as a scaffold for inhibition of the Tissue Factor (TF)/Factor VIIa (FVIIa). In contrast to tissue factor pathway inhibitor (TFPI), however, Ixolaris does not bind to the active site cleft of FXa. Instead, complex formation is mediated by the FXa heparin-binding exosite. Due to its potent and long-lasting antithrombotic activity, Ixolaris is a promising agent for anticoagulant therapy...
November 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28685146/his-flag-tag-as-a-fusion-partner-of-glycosylated-human-interferon-gamma-and-its-mutant-gain-or-loss
#12
Elena Krachmarova, Milena Tileva, Elena Lilkova, Peicho Petkov, Klaus Maskos, Nevena Ilieva, Ivan Ivanov, Leandar Litov, Genoveva Nacheva
In order to obtain glycosylated human interferon-gamma (hIFNγ) and its highly prone to aggregation mutant K88Q, a secretory expression in insect cells was employed. To facilitate recombinant proteins purification, detection, and stability the baculovirus expression vectors were constructed to bear N-terminal His6-FLAG tag. Although the obtained proteins were glycosylated, we found that their biological activity was 100 times lower than expected. Our attempts to recover the biological properties of both proteins by tag removal failed due to enterokinase resistance of the tag...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28666856/production-of-recombinant-salmon-insulin-like-growth-factor-binding-protein-1-subtypes
#13
Hanae Tanaka, Gakuto Oishi, Yusuke Nakano, Hiroko Mizuta, Yuta Nagano, Naoshi Hiramatsu, Hironori Ando, Munetaka Shimizu
Insulin-like growth factor (IGF)-I is a growth promoting hormone that exerts its actions through endocrine, paracrine and autocrine modes. Local IGF-I is essential for normal growth, whereas circulating IGF-I plays a crucial role in regulating the production and secretion of growth hormone (GH) by the pituitary gland. These actions of IGF-I are modulated by six insulin-like growth factor binding proteins (IGFBPs). In teleosts, two subtypes of each IGFBP are present due to an extra round of whole-genome duplication...
June 27, 2017: General and Comparative Endocrinology
https://www.readbyqxmd.com/read/28586021/construction-and-characterization-of-a-transmembrane-eukaryotic-expression-vector-based-on-the-membrane-domain-structure-of-tnf-%C3%AE
#14
Fa Wang, Liang Zeng, Yue-Li Wang, Shi-Quan Cui, Liang Hu, Jun-Ming Zheng, Di-Nan Huang, Gan Hou
The aim of the present study was to construct a fast-acting, eukaryotic expression vector in eukaryotic cells based on transmembrane-tumor necrosis factor‑α (TM‑TNF‑α) structure. Two types of recombinant eukaryotic expression vectors were constructed, pcDNA3.1-TM-enterokinase-TNF‑α and pcDNA3.1‑TM‑Factor Xa‑TNF‑α, according to the TNF‑α transmembrane segments. Following the generation of these vectors, mouse embryonic 3T3 fibroblasts were transfected and reverse transcription‑polymerase chain reaction and western blotting analyses were used to analyze mTNF‑α mRNA and protein expression levels, respectively, in total cellular protein extracts and extracellular fluid...
August 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28510446/modularly-constructed-synthetic-granzyme-b-molecule-enables-interrogation-of-intracellular-proteases-for-targeted-cytotoxicity
#15
Patrick Ho, Christopher Ede, Yvonne Y Chen
Targeted therapies promise to increase the safety and efficacy of treatments against diseases ranging from cancer to viral infections. However, the vast majority of targeted therapeutics relies on the recognition of extracellular biomarkers, which are rarely restricted to diseased cells and are thus prone to severe and sometimes-fatal off-target toxicities. In contrast, intracellular antigens present a diverse yet underutilized repertoire of disease markers. Here, we report a protein-based therapeutic platform-termed Cytoplasmic Oncoprotein VErifier and Response Trigger (COVERT)-which enables the interrogation of intracellular proteases to trigger targeted cytotoxicity...
August 18, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28218499/fine-tuning-sortase-mediated-immobilization-of-protein-layers-on-surfaces-using-sequential-deprotection-and-coupling
#16
Maryam Raeeszadeh-Sarmazdeh, Ranganath Parthasarathy, Eric T Boder
Increasing interest in protein immobilization on surfaces has heightened the need for techniques enabling layer-by-layer protein attachment. Here, we report a technique for controlling enzyme-mediated immobilization of layers of protein on the surface using a genetically encoded protecting group. An enterokinase-cleavable peptide sequence was inserted at the N-terminus of bifunctional fluorescent proteins containing Sortase A substrate recognition tags at both ends to control Sortase A-mediated protein immobilization on the surface layer-by-layer...
February 20, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28159581/recombinant-expression-of-intrepicalcin-from-the-scorpion-vaejovis-intrepidus-and-its-effect-on-skeletal-ryanodine-receptors
#17
Leonel Vargas-Jaimes, Liang Xiao, Jing Zhang, Lourival D Possani, Héctor H Valdivia, Verónica Quintero-Hernández
BACKGROUND: Scorpion venoms contain toxins that modulate ionic channels, among which are the calcins, a small group of short, basic peptides with an Inhibitor Cystine Knot (ICK) motif that target calcium release channels/ryanodine receptors (RyRs) with high affinity and selectivity. Here we describe the heterologous expression of Intrepicalcin, identified by transcriptomic analysis of venomous glands from Vaejovis intrepidus. METHODS: Recombinant Intrepicalcin was obtained in Escherichia coli BL21-DE3 (periplasm) by fusing the Intrepicalcin gene to sequences coding for signal-peptide, thioredoxin, His-tag and enterokinase cleavage site...
April 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28126552/design-and-expression-of-recombinant-toxins-from-mexican-scorpions-of-the-genus-centruroides-for-production-of-antivenoms
#18
J M Jiménez-Vargas, V Quintero-Hernández, L González-Morales, E Ortiz, L D Possani
This manuscript describes the design of plasmids containing the genes coding for four main mammalian toxins of scorpions from the genus Centruroides (C.) of Mexico. The genes that code for toxin 2 of C. noxius (Cn2), toxin 2 from C. suffusus (Css2) and toxins 1 and 2 from C. limpidus (Cll1 and Cll2) were included into individual plasmids carrying the genetic construction for expression of fusion proteins containing a leader peptide (pelB) that directs the expressed protein to the bacterial periplasm, a carrier protein (thioredoxin), the cleavage site for enterokinase, the chosen toxin and a poly-histidine tag (6xHis-tag) for purification of the hybrid protein by immobilized metal ion affinity chromatography after expression in Escherichia coli strain BL21 (DE3)...
March 15, 2017: Toxicon: Official Journal of the International Society on Toxinology
https://www.readbyqxmd.com/read/28075401/anti-neuroblastoma-properties-of-a-recombinant-sunflower-lectin
#19
Marcela Pinedo, Melanie Genoula, María Ximena Silveyra, André De Oliveira Carvalho, Mariana Regente, Marianela Del Río, Júlia Ribeiro Soares, Valdirene Moreira Gomes, Laura De La Canal
According to their sugar recognition specificity, plant lectins are proposed as bioactive proteins with potential in cancer treatment and diagnosis. Helja is a mannose-specific jacalin-like lectin from sunflower which was shown to inhibit the growth of certain fungi. Here, we report its recombinant expression in a prokaryotic system and its activity in neurobalstoma cells. Helja coding sequence was fused to the pET-32 EK/LIC, the enterokinase/Ligation-independent cloning vector and a 35 kDa protein was obtained in Escherichia coli representing Helja coupled to thioredoxin (Trx)...
January 10, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27977898/expansion-of-divergent-sea-domains-in-cell-surface-proteins-and-nucleoporin-54
#20
Jimin Pei, Nick V Grishin
SEA (sea urchin sperm protein, enterokinase, agrin) domains, many of which possess autoproteolysis activity, have been found in a number of cell surface and secreted proteins. Despite high sequence divergence, SEA domains were also proposed to be present in dystroglycan based on a conserved autoproteolysis motif and receptor-type protein phosphatase IA-2 based on structural similarity. The presence of a SEA domain adjacent to the transmembrane segment appears to be a recurring theme in quite a number of type I transmembrane proteins on the cell surface, such as MUC1, dystroglycan, IA-2, and Notch receptors...
March 2017: Protein Science: a Publication of the Protein Society
keyword
keyword
37615
1
2
Fetch more papers »
Fetching more papers... Fetching...
Read by QxMD. Sign in or create an account to discover new knowledge that matter to you.
Remove bar
Read by QxMD icon Read
×

Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"