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Protein ligation

Marici R Rached, Verônica Coelho, Maria Lúcia C Marin, Kátia Pincerato, André Fujita, Jorge E Kalil, Maurício S Abrão
OBJECTIVE: To assess whether the HLA-G immunomodulatory protein is potentially involved in the pathophysiology of endometriosis or disease progression. STUDY DESIGN: Cross-sectional observational study of 227 women who underwent laparoscopy, being 146 for endometriosis excision and 81 for elective tubal ligation (control group). Soluble HLA-G (sHLA-G) levels in the serum and peritoneal fluid (PF), as well as the HLA-G protein expression in matched eutopic and ectopic endometrium of women with and without endometriosis were evaluated by ELISA and immunohistochemistry assays, respectively...
February 6, 2019: European Journal of Obstetrics, Gynecology, and Reproductive Biology
Eiji Nakata, Huyen Dinh, Thang Minh Nguyen, Takashi Morii
DNA nanostructures serve as the ideal scaffolds to assemble materials of interest. Among these, proteins are of particularly interesting class of molecules to assemble because of their huge functional variability. Sequence-specific DNA binding proteins have been applied as adaptors to stably locate the fused proteins at defined positions of DNA scaffold in high loading yields. The strategy allows to control the number of enzyme molecules and to maintain the catalytic activity. By fusing a chemoselective self-ligating protein tag to the DNA binding protein, the modular adaptors formed covalent bonds at respective sequences on DNA scaffold with fast reaction kinetics...
2019: Methods in Enzymology
Weixin Sun, Renyou Pan, Jun Song, Honglin Sun
Effect of simvastatin on the expression of caspase-3 in myocardial ischemia reperfusion injury in rats was observed to explore the protective effect of caspase-3 through anti-apoptosis mechanism. A total of 48 healthy male SD rats weighing 160-240 g were selected and divided into 4 groups randomly, namely, the blank group, the sham operation group, the ischemia-reperfusion group and the simvastatin group, with 12 rats in each group. The model of SD rats was made by ligation. The loosen ligature made the reperfusion animal model, the occurrence of arrhythmia in the electrocardiogram of lead II in the experimental animal model was observed, and the area of myocardial infarction in the experimental animal models was detected...
March 2019: Experimental and Therapeutic Medicine
Do Young Hyeon, Jong Hyun Kim, Tae Jin Ahn, Yeshin Cho, Daehee Hwang, Sunghoon Kim
Aminoacyl-tRNA synthetases (ARSs) are enzymes that ligate their cognate amino acids to tRNAs for protein synthesis. However, recent studies have shown that their functions are expanded beyond protein synthesis through the interactions with diverse cellular factors. In this review, we discuss how ARSs have evolved to expand and control their functions by forming protein assemblies. We particularly focus on a macromolecular ARS complex in eukaryotes, named multi-tRNA synthetase complex (MSC), which is proposed to provide a channel through which tRNAs reach bound ARSs to receive their cognate amino acid and transit further to the translation machinery...
February 19, 2019: Journal of Biological Chemistry
Zeyu Li, Yifan Jia, Yang Feng, Ruixia Cui, Runchen Miao, Xing Zhang, Kai Qu, Chang Liu, Jingyao Zhang
Sepsis is defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection. Methane has been reported to have anti-oxidative, anti-apoptotic and anti-inflammatory properties. We investigated the potential protective effects of methane on sepsis-induced injury and determined the related mechanisms. C57BL/6 mice received laparotomy with cecal ligation and puncture (CLP) to create a septic model, followed by methane-rich saline (MRS) treatment after CLP. MRS treatment improved the 5-day survival rate and organ functions and alleviated pathological damage of the mice, as well as reduced excessive inflammatory mediators, such as tumor necrosis factor-α and interleukin-6...
February 18, 2019: Aging
Y Zhou, D-D Peng, H Chong, S-Q Zheng, F Zhu, G Wang
OBJECTIVE: To investigate the effect of isoflurane on myocardial ischemia-reperfusion injury through the p38 mitogen-activated protein kinase (MAPK) signaling pathway. MATERIALS AND METHODS: A total of 36 specific-pathogen-free (SPF) Sprague-Dawley rats were randomly divided into sham group (n=12), model group (n=12) and isoflurane group (n=12). In model group and isoflurane group, the myocardial ischemia-reperfusion injury model was established via the ligation of left anterior descending coronary artery (ischemia for 30 min and reperfusion for 3 h)...
February 2019: European Review for Medical and Pharmacological Sciences
Ryan M Young
Many methods exist to study oncogenic signaling in lymphoma cells, each with certain strengths and weaknesses. The proximity ligation assay (PLA) is a technique used to examine protein-protein interactions in situ. PLA offers many advantages over traditional biochemical approaches to signaling in that it can detect both transient interactions, which are hard to capture otherwise, as well as protein interactions within large supramolecular signaling platforms that are often intractable to traditional biochemical methodologies...
2019: Methods in Molecular Biology
Bipandeep Dhillon, Fatemah Aleithan, Zahi Abdul-Sater, Ali A Abdul-Sater
TRAFs [tumor necrosis factor (TNF) receptor associated factors] are a family of signaling molecules that function downstream of multiple receptor signaling pathways and play a pivotal role in the biology of innate, and adaptive immune cells. Following receptor ligation, TRAFs generally function as adapter proteins to mediate the activation of intracellular signaling cascades. With the exception of TRAF1 that lacks a Ring domain, TRAFs have an E3 ubiquitin ligase activity which also contributes to their ability to activate downstream signaling pathways...
2019: Frontiers in Immunology
Meizhen Zheng, Simon Zhongyuan Tian, Daniel Capurso, Minji Kim, Rahul Maurya, Byoungkoo Lee, Emaly Piecuch, Liang Gong, Jacqueline Jufen Zhu, Zhihui Li, Chee Hong Wong, Chew Yee Ngan, Ping Wang, Xiaoan Ruan, Chia-Lin Wei, Yijun Ruan
The genomes of multicellular organisms are extensively folded into 3D chromosome territories within the nucleus1 . Advanced 3D genome-mapping methods that combine proximity ligation and high-throughput sequencing (such as chromosome conformation capture, Hi-C)2 , and chromatin immunoprecipitation techniques (such as chromatin interaction analysis by paired-end tag sequencing, ChIA-PET)3 , have revealed topologically associating domains4 with frequent chromatin contacts, and have identified chromatin loops mediated by specific protein factors for insulation and regulation of transcription5-7 ...
February 18, 2019: Nature
Li Zhang, Zhihong Yang, Wendong Huang, Jianguo Wu
Cholestasis induces the hepatic long non-coding RNA H19, which promotes the progression of cholestatic liver fibrosis. However, microRNAs that are dysregulated by H19 during cholestasis remain elusive. Using miRNA-sequencing analysis followed by qPCR validation, we identified marked upregulation of eight members of the let-7 family in cholestatic livers by bile duct ligation (BDL) and H19 overexpression. In particular, the expression of let-7a-1/7d/7f-1 was highly induced in H19-BDL livers but decreased in H19KO-BDL livers...
February 18, 2019: Cell Death & Disease
Tingting Yu, Xin Zhang, Haosong Shi, Jinge Tian, Lingling Sun, Xueming Hu, Wenqiang Cui, Dongping Du
Peripheral nerve injury causes neuropathic pain and microglia activation. P2Y12 receptors on microglia are thought to be a key player in the surveillance of the local environment, but whether or how these receptors are engaged in the cross-talk between microglia and neurons of the dorsal horn remain ambiguous. Using a rodent model of nerve injury-induced pain, we investigated the roles of P2Y12 in microglia activation, excitatory synaptic transmission, and nociceptive allodynia. We found that spinal nerve ligation (SNL) significantly increased the level of P2Y12 receptors specifically in the microglia of the ipsilateral dorsal horn...
February 18, 2019: Cell Death & Disease
Teppei Kitano, Soichiro Usui, Shin-Ichiro Takashima, Oto Inoue, Chiaki Goten, Ayano Nomura, Kazuaki Yoshioka, Masaki Okajima, Shuichi Kaneko, Yoh Takuwa, Masayuki Takamura
Vascular remodeling, resulting from proliferation and migration of vascular smooth muscle cells (VSMCs), is a major cause of atherosclerosis and restenosis. The lysophospholipid mediator sphingosine-1-phosphate (S1P) regulates proliferation and migration of VSMCs via S1P-specific G protein-coupled receptors, including S1P receptor 1 (S1PR1) to S1PR3. However, the role of S1PR1 in vascular remodeling is not well understood. Therefore, in this study, we aimed to investigate the effect of S1PR1 on neointimal hyperplasia in a carotid artery ligation mouse model using transgenic C57Bl/6 mice that overexpressed S1PR1 (Tg-S1PR1) under the control of α-smooth muscle actin promoter...
February 15, 2019: Biochemical and Biophysical Research Communications
Darja Pollpeter, Andrew Sobala, Michael H Malim
Monitoring of nucleic acid intermediates during virus replication provides insights into the effects and mechanisms of action of antiviral compounds and host cell proteins on viral DNA synthesis. Here we address the lack of a cell-based, high-coverage, and high-resolution assay that is capable of defining retroviral reverse transcription intermediates within the physiological context of virus infection. The described method captures the 3'-termini of nascent complementary DNA (cDNA) molecules within HIV-1 infected cells at single nucleotide resolution...
January 30, 2019: Journal of Visualized Experiments: JoVE
Wen Wu, Wei Zhang, Mihyun Choi, Jinjing Zhao, Ping Gao, Min Xue, Harold A Singer, David Jourd'heuil, Xiaochun Long
Injury-induced stenosis is a serious vascular complication. We previously reported that p38α (MAPK14), a redox-regulated p38MAPK family member was a negative regulator of the VSMC contractile phenotype in vitro. Here we evaluated the function of VSMC-MAPK14 in vivo in injury-induced neointima hyperplasia and the underlying mechanism using an inducible SMC-MAPK14 knockout mouse line (iSMC-MAPK14-/- ). We show that MAPK14 expression and activity were induced in VSMCs after carotid artery ligation injury in mice and ex vivo cultured human saphenous veins...
February 6, 2019: Redox Biology
Yina Liu, Shengtian Mu, Xu Li, Yingjian Liang, Liang Wang, Xiaochun Ma
BACKGROUND: Unfractionated heparin (UFH) has been shown to ameliorate lung edema and lung vascular leakage in lipopolysaccharide-induced lung injury. Impaired tight junction (TJ) function is a sign of sepsis-induced acute respiratory distress syndrome (ARDS) and acute lung injury (ALI), which is closely related to the downregulated expression of TJ-specific proteins or the upregulated expression of inflammatory cytokines. Because UFH has been intensively studied in modulating inflammation, we hypothesize that UFH may play a positive role in treating sepsis-induced ARDS/ALI by protecting TJs...
February 13, 2019: Journal of Surgical Research
Lévêque Romain, Corbet Cyril, Aubert Léo, Guilbert Matthieu, Lagadec Chann, Adriaenssens Eric, Duval Jérémy, Finetti Pascal, Birnbaum Daniel, Magné Nicolas, Chopin Valérie, Bertucci François, Le Bourhis Xuefen, Toillon Robert-Alain
ProNGF expression has been linked to several types of cancers including breast cancer, and we have previously shown that proNGF stimulates breast cancer invasion in an autocrine manner through membrane receptors sortilin and TrkA. However, little is known regarding TrkA-associated protein partners upon proNGF stimulation. By proteomic analysis and proximity ligation assays, we found that proNGF binding to sortilin induced sequential formation of the functional sortilin/TrkA/EphA2 complex, leading to TrkA-phosphorylation dependent Akt activation and EphA2-dependent Src activation...
February 13, 2019: Cancer Letters
Mei Yee Kwan, Anthony Choo, Taleen Hanania, Afshin Ghavami, Jose Beltran, John Shea, Amidi Barboza, Andrew Hu, Marcie Fowler, Venugopal Rao Neelagiri, Irving Sucholeiki
There is an unmet medical need for the development of non-addicting pain therapeutics with enhanced efficacy and tolerability. The current study examined the effects of AQU-118, an orally active inhibitor of metalloproteinase-2 (MMP-2) and MMP-9, in the spinal nerve ligation (SNL) rat model of neuropathic pain. Mechanical allodynia and the levels of various biomarkers were examined within the dorsal root ganglion (DRG) before and after oral dosing with AQU-118. The rats that received the SNL surgery exhibited significant mechanical allodynia as compared to sham controls...
February 14, 2019: International Journal of Molecular Sciences
Adelaide Tousley, Maria Iuliano, Elizabeth Weisman, Ellen Sapp, Heather Richardson, Petr Vodicka, Jonathan Alexander, Neil Aronin, Marian DiFiglia, Kimberly B Kegel-Gleason
One response of cells to growth factor stimulus involves changes in morphology driven by the actin cytoskeleton and actin associated proteins which regulate functions such as cell adhesion, motility and in neurons, synaptic plasticity. Previous studies suggest that Huntingtin may be involved in regulating morphology however, there has been limited evidence linking endogenous Huntingtin localization or function with cytoplasmic actin in cells. We found that depletion of Huntingtin in human fibroblasts reduced adhesion and altered morphology and these phenotypes were made worse with growth factor stimulation, whereas the presence of the Huntington's Disease mutation inhibited growth factor induced changes in morphology and increased numbers of vinculin-positive focal adhesions...
2019: PloS One
Qiao-Mei Xie, Qiu-Yue Lou, Shun-Wei Huang, Hua-Qing Hu, Su-Su Li, Man Zhang, Xiu-Xiu Sun, Jian-Hua Xu, Shan-Qun Jiang, Sheng-Xiu Liu, Sheng-Qian Xu, Jing Cai, Shuang Liu, Fa-Ming Pan, Jin-Hui Tao, Long Qian, Chun-Huai Wang, Chun-Mei Liang, Hai-Liang Huang, Hai-Feng Pan, Hong Su, Yan-Feng Zou
OBJECTIVES: The aim of this study is to investigate whether heat shock protein 70 (Hsp70) gene polymorphisms are implicated in systemic lupus erythematous (SLE) susceptibility, the efficacy of glucocorticoids (GCs) treatment, and improvement of health-related quality of life. METHODS: A total of 499 SLE patients and 499 controls were included in a case-control study, and 468 SLE patients treated with GCs for 12 weeks were involved in a follow-up study. Patients who completed the 12-week follow-up were divided into GCs-sensitive and GCs-insensitive group by using the SLE disease activity index...
February 13, 2019: Journal of Clinical Rheumatology: Practical Reports on Rheumatic & Musculoskeletal Diseases
Gang Li, Raymond Moellering
Synthesis of antibody-oligonucleotide conjugates has enabled the development of highly sensitive bioassays for specific epitopes in the laboratory and clinic. Most synthetic schemes to generate these hybrid molecules require expensive reagents, significant quantities of input antibody, and multi-step purification routes, thus limiting widespread application. Here we report a facile and robust conjugation strategy that involves "plug-and-play" antibody conjugation with succinimidyl-functionalized oligonucleotides, which are high-yielding and compatible for use directly after buffer exchange...
February 14, 2019: Chembiochem: a European Journal of Chemical Biology
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