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https://www.readbyqxmd.com/read/29115621/effects-of-shield1-on-the-viral-replication-of-varicella%C3%A2-zoster-virus-containing-fkbp%C3%A2-tagged-orf4-and-48
#1
Shuying Li, Zhanjun Liu, Ji Li, Aihua Liu, Lihua Zhu, Kui Yu, Ke Zhang
The present study aimed to explore the effects of a stabilizing ligand, Shield‑1, on the replication of recombinant varicella‑zoster virus (VZV) containing FK506 binding protein (FKPB) tags in essential open reading frames (ORF) 4 and 48. A specific galactokinase (galK) selection method was conducted, following the addition of galK labels to VZV ORF4 and 48, using a SW102 VZV bacterial artificial chromosome (BAC) system. Subsequently, recombinant VZV containing FKPB tags in ORF4 and 48 was constructed by counterselection and homologous recombination...
November 6, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29109782/berunda-polypeptides-multi-headed-fusion-proteins-promote-subcutaneous-administration-of-rapamycin-to-breast-cancer-in-vivo
#2
Jugal P Dhandhukia, Zhe Li, Santosh Peddi, Shruti Kakan, Arjun Mehta, David Tyrpak, Jordan Despanie, J Andrew MacKay
Recombinant Elastin-Like Polypeptides (ELPs) serve as attractive scaffolds for nanoformulations because they can be charge-neutral, water soluble, high molecular weight, monodisperse, biodegradable, and decorated with functional proteins. We recently reported that fusion of the FK-506 binding protein 12 (FKBP) to an ELP nanoparticle (FSI) reduces rapamycin (Rapa) toxicity and enables intravenous (IV) therapy in both a xenograft breast cancer model and a murine autoimmune disease model. Rapa has poor solubility, which leads to variable oral bioavailability or drug precipitation following parenteral administration...
2017: Theranostics
https://www.readbyqxmd.com/read/29073761/targeted-gene-repression-using-novel-bifunctional-molecules-to-harness-endogenous-histone-deacetylation-activity
#3
Kyle V Butler, Anna M Chiarella, Jian Jin, Nathaniel A Hathaway
Epigenome editing is a powerful method for life science research and could give rise to new therapies for diseases initiated or maintained by epigenetic dysregulation, including several types of cancers and autoimmune disorders. In addition, much is still unknown about the mechanisms by which histone-modifying proteins work in concert to properly regulate gene expression. To investigate and manipulate complex epigenetic interactions in live cells, we have developed a small molecule platform for specifically inducing gene repression and histone deacetylation at a reporter gene...
November 7, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/29042512/in-vitro-reconstitution-of-t-cell-receptor-mediated-segregation-of-the-cd45-phosphatase
#4
Catherine B Carbone, Nadja Kern, Ricardo A Fernandes, Enfu Hui, Xiaolei Su, K Christopher Garcia, Ronald D Vale
T cell signaling initiates upon the binding of peptide-loaded MHC (pMHC) on an antigen-presenting cell to the T cell receptor (TCR) on a T cell. TCR phosphorylation in response to pMHC binding is accompanied by segregation of the transmembrane phosphatase CD45 away from TCR-pMHC complexes. The kinetic segregation hypothesis proposes that CD45 exclusion shifts the local kinase-phosphatase balance to favor TCR phosphorylation. Spatial partitioning may arise from the size difference between the large CD45 extracellular domain and the smaller TCR-pMHC complex, although parsing potential contributions of extracellular protein size, actin activity, and lipid domains is difficult in living cells...
October 31, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29036638/the-basic-tilted-helix-bundle-domain-of-the-prolyl-isomerase-fkbp25-is-a-novel-double-stranded-rna-binding-module
#5
David Dilworth, Santosh K Upadhyay, Pierre Bonnafous, Amiirah Bibi Edoo, Sarah Bourbigot, Francy Pesek-Jardim, Geoff Gudavicius, Jason J Serpa, Evgeniy V Petrotchenko, Christoph H Borchers, Christopher J Nelson, Cameron D Mackereth
Prolyl isomerases are defined by a catalytic domain that facilitates the cis-trans interconversion of proline residues. In most cases, additional domains in these enzymes add important biological function, including recruitment to a set of protein substrates. Here, we report that the N-terminal basic tilted helix bundle (BTHB) domain of the human prolyl isomerase FKBP25 confers specific binding to double-stranded RNA (dsRNA). This binding is selective over DNA as well as single-stranded oligonucleotides. We find that FKBP25 RNA-association is required for its nucleolar localization and for the vast majority of its protein interactions, including those with 60S pre-ribosome and early ribosome biogenesis factors...
September 25, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29032102/regulation-of-ca-2-signaling-by-acute-hypoxia-and-acidosis-in-rat-neonatal-cardiomyocytes
#6
José-Carlos Fernández-Morales, Martin Morad
Ischemic heart disease is an arrhythmogenic condition, accompanied by hypoxia, acidosis, and impaired Ca(2+) signaling. Here we report on effects of acute hypoxia and acidification in rat neonatal cardiomyocytes cultures. RESULTS: Two populations of neonatal cardiomyocyte were identified based on inactivation kinetics of L-type ICa: rapidly-inactivating ICa (τ~20ms) myocytes (prevalent in 3-4-day cultures), and slow-inactivating ICa (τ≥40ms) myocytes (dominant in 7-day cultures). Acute hypoxia (pO2<5mmHg for 50-100s) suppressed ICa reversibly in both cell-types to different extent and with different kinetics...
October 12, 2017: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/29024217/use-of-the-target-of-rapamycin-fatc-domain-as-a-general-membrane-anchor-the-fkbp-12-like-domain-of-fkbp38-as-a-case-study
#7
Maristella De Cicco, Lech-G Milroy, Sonja A Dames
Increased efforts have been undertaken to better understand the formation of signaling complexes at cellular membranes. Since the preparation of proteins containing a transmembrane domain or a prenylation motif is generally challenging an alternative membrane anchoring unit that is easy to attach, water-soluble and binds to different membrane mimetics would find broad application. The 33-residue long FATC domain of yeast TOR1 (y1fatc) fulfills these criteria and binds to neutral and negatively charged micelles, bicelles and liposomes...
October 12, 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28982713/publisher-s-note-association-of-fk506-binding-proteins-with-ryr-channels-effect-of-clic2-binding-on-sub-conductance-opening-and-fkbp-binding-j-cell-sci-doi-10-1242-jcs-204461
#8
Spencer J Richardson, Gregory A Steele, Esther M Gallant, Alexander Lam, Charles E Schwartz, Philip G Board, Marco G Casarotto, Nicole A Beard, Angela F Dulhunty
No abstract text is available yet for this article.
October 5, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28973376/the-integrity-and-organization-of-the-human-aipl1-functional-domains-is-critical-for-its-role-as-a-hsp90-dependent-co-chaperone-for-rod-pde6
#9
Almudena Sacristan-Reviriego, James Bellingham, Chrisostomos Prodromou, Neruban Kumaran, James Bainbridge, Michel Michaelides, Jacqueline van der Spuy
Biallelic mutations in the photoreceptor-expressed aryl hydrocarbon receptor interacting protein-like 1 (AIPL1) are associated with autosomal recessive Leber congenital amaurosis (LCA), the most severe form of inherited retinopathy in early childhood. AIPL1 functions as a photoreceptor-specific co-chaperone that interacts with the molecular chaperone HSP90 to facilitate the stable assembly of the retinal cyclic GMP (cGMP) phosphodiesterase (PDE6) holoenzyme. In this study, we characterized the functional deficits of AIPL1 variations, some of which induce aberrant pre-mRNA AIPL1 splicing leading to the production of alternative AIPL1 isoforms...
November 15, 2017: Human Molecular Genetics
https://www.readbyqxmd.com/read/28965803/expression-purification-and-characterization-of-a-catalytic-domain-of-human-protein-tyrosine-phosphatase-non-receptor-12-ptpn12-in-escherichia-coli-with-fkbp-type-ppiase-as-a-chaperon
#10
Yuan Sui, Xingye Fu, Yuchen Wang, Weiyan Hu, Tong Zhang, Wanyao Liu, Liyan Jiang, Shu Xing, Xueqi Fu, Xuesong Xu
Protein tyrosine phosphatase non-receptor type 12 (PTPN12), also known as PTP-PEST, was broadly expressed in hemopoietic cells. Recent research has shown that this enzyme is involved in tumorigenesis, as well as in tumor progression and transfer, as it can suppress multiple oncogenic tyrosine kinases. However, the difficulty of soluble expression of PTP-PEST in prokaryotic cells has resulted in great limitations in investigating its structure and functions. In this study, we successfully carried out soluble expression of the catalytic domain of PTP-PEST (ΔPTP-PEST) in Escherichia coli and performed an enzymatic characterization and kinetics...
September 28, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28955036/optimizing-the-fragment-complementation-of-apex2-for-detection-of-specific-protein-protein-interactions-in-live-cells
#11
Miaomiao Xue, Junjie Hou, Linlin Wang, Dongwan Cheng, Jingze Lu, Li Zheng, Tao Xu
Dynamic protein-protein interactions (PPIs) play crucial roles in cell physiological processes. The protein-fragment complementation (PFC) assay has been developed as a powerful approach for the detection of PPIs, but its potential for identifying protein interacting regions is not optimized. Recently, an ascorbate peroxidase (APEX2)-based proximity-tagging method combined with mass spectrometry was developed to identify potential protein interactions in live cells. In this study, we tested whether APEX2 could be employed for PFC...
September 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28937754/bifunctional-elastin-like-polypeptide-nanoparticles-bind-rapamycin-and-integrins-and-suppress-tumor-growth-in-vivo
#12
Jugal P Dhandhukia, Pu Shi, Santosh Peddi, Zhe Li, Suhaas Aluri, Yaping Ju, Dab Brill, Wan Wang, Siti M Janib, Yi-An Lin, Shuanglong Liu, Honggang Cui, J Andrew MacKay
Recombinant protein-polymer scaffolds such as elastin-like polypeptides (ELPs) offer drug-delivery opportunities including biocompatibility, monodispersity, and multifunctionality. We recently reported that the fusion of FK-506 binding protein 12 (FKBP) to an ELP nanoparticle (FSI) increases rapamycin (Rapa) solubility, suppresses tumor growth in breast cancer xenografts, and reduces side effects observed with free-drug controls. This new report significantly advances this carrier strategy by demonstrating the coassembly of two different ELP diblock copolymers containing drug-loading and tumor-targeting domains...
October 12, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28916764/rapid-and-reversible-epigenome-editing-by-endogenous-chromatin-regulators
#13
Simon M G Braun, Jacob G Kirkland, Emma J Chory, Dylan Husmann, Joseph P Calarco, Gerald R Crabtree
Understanding the causal link between epigenetic marks and gene regulation remains a central question in chromatin biology. To edit the epigenome we developed the FIRE-Cas9 system for rapid and reversible recruitment of endogenous chromatin regulators to specific genomic loci. We enhanced the dCas9-MS2 anchor for genome targeting with Fkbp/Frb dimerizing fusion proteins to allow chemical-induced proximity of a desired chromatin regulator. We find that mSWI/SNF (BAF) complex recruitment is sufficient to oppose Polycomb within minutes, leading to activation of bivalent gene transcription in mouse embryonic stem cells...
September 15, 2017: Nature Communications
https://www.readbyqxmd.com/read/28851804/fkbp-association-with-ryr-channels-effect-of-clic2-binding-on-sub-conductance-opening-and-fkbp-binding
#14
Spencer J Richardson, Gregory A Steele, Esther M Gallant, Alexander Lam, Charles E Schwartz, Philip G Board, Marco G Casarotto, Nicole A Beard, Angela F Dulhunty
Ryanodine receptor (RyR) calcium channels are central to striated muscle function and influence signalling in neurones and other cell types. Beneficially low RyR activity and maximum conductance opening may be stabilised when RyRs bind to FK506 binding proteins (FKBPs) and destabilised by FKBP dissociation, with submaximal opening during RyR hyperactivity associated with myopathies and neurological disorders. However this is debated and quantitative evidence is lacking. Here we have measured altered FKBP binding to RyRs and submaximal activity with addition of wild-type (WT) CLIC2, an inhibitory RyR ligand, or its H101Q mutant that hyperactivates RyRs, likely causing cardiac and intellectual abnormalities...
August 29, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28807154/total-internal-reflectance-fluorescence-imaging-of-genetically-engineered-ryanodine-receptor-targeted-ca-2-probes-in-rat-ventricular-myocytes
#15
Sara Pahlavan, Marin Morad
The details of cardiac Ca(2+) signaling within the dyadic junction remain unclear because of limitations in rapid spatial imaging techniques, and availability of Ca(2+) probes localized to dyadic junctions. To critically monitor ryanodine receptors' (RyR2) Ca(2+) nano-domains, we combined the use of genetically engineered RyR2-targeted pericam probes, (FKBP-YCaMP, Kd=150nM, or FKBP-GCaMP6, Kd=240nM) with rapid total internal reflectance fluorescence (TIRF) microscopy (resolution, ∼80nm). The punctate z-line patterns of FKBP,(2)-targeted probes overlapped those of RyR2 antibodies and sharply contrasted to the images of probes targeted to sarcoplasmic reticulum (SERCA2a/PLB), or cytosolic Fluo-4 images...
September 2017: Cell Calcium
https://www.readbyqxmd.com/read/28668496/generation-of-nanobodies-against-slyd-and-development-of-tools-to-eliminate-this-bacterial-contaminant-from-recombinant-proteins
#16
Yaozhong Hu, Ema Romão, Didier Vertommen, Cécile Vincke, Francisco Morales-Yánez, Carlos Gutiérrez, Changxiao Liu, Serge Muyldermans
The gene for a protein domain, derived from a tumor marker, fused to His tag codons and under control of a T7 promotor was expressed in E. coli strain BL21 (DE3). The recombinant protein was purified from cell lysates through immobilized metal affinity chromatography and size-exclusion chromatography. A contaminating bacterial protein was consistently co-purified, even using stringent washing solutions containing 50 or 100 mM imidazole. Immunization of a dromedary with this contaminated protein preparation, and the subsequent generation and panning of the immune Nanobody library yielded several Nanobodies of which 2/3 were directed against the bacterial contaminant, reflecting the immunodominance of this protein to steer the dromedary immune response...
September 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28652145/diverse-structures-functions-and-uses-of-fk506-binding-proteins
#17
REVIEW
Julia Maeve Bonner, Gabrielle L Boulianne
FK506 (Tacrolimus), isolated from Streptomyces tsukubaenis is a powerful immunosuppressant shown to inhibit T cell activation. FK506 mediated immunosuppression requires the formation of a complex between FK506, a FK506 binding protein (FKBP) and calcineurin. Numerous FKBPs have been identified in a wide range of species, from single celled organisms to humans. FKBPs show peptidylprolyl cis/trans isomerase (PPIase) activity and have been shown to affect a wide range of cellular processes including protein folding, receptor signaling and apoptosis...
June 23, 2017: Cellular Signalling
https://www.readbyqxmd.com/read/28636428/fk506-binding-proteins-12-and-12-6-fkbps-as-regulators-of-cardiac-ryanodine-receptors-insights-from-new-functional-and-structural-knowledge
#18
Luis A Gonano, Peter P Jones
Ryanodine Receptors (RyRs) are intracellular Ca(2+) channels that mediate Ca(2+) flux from the sarco(endo)plasmic reticulum in many cell types. The interaction of RyRs with FK506-binding proteins (FKBPs) has been proposed as an important regulatory mechanism, where the loss of this interaction leads to channel dysfunction. In the heart, phosphorylation of RyR has been suggested to disrupt the RyR-FKBP interaction promoting altered Ca(2+) signaling, heart failure and arrhythmias. However, the functional result of FKBP interaction with RyR and how this interaction is regulated remains highly controversial...
September 3, 2017: Channels
https://www.readbyqxmd.com/read/28631479/quantitative-analysis-of-ligand-induced-heterodimerization-of-two-distinct-receptors
#19
Chang Lu, Zhi-Xin Wang
The induced dimerization of two distinct receptors through a heterobifunctional inducer is prevalent among all levels of cellular signaling processes, yet its complexity poses difficulty for systematic quantitative analysis. This paper first shows how to calculate the amount of any possible complex or monomer of heteroligand and two receptors present at equilibrium. The theory is subsequently applied to the determination of three independent equilibrium parameters involved in the rapamycin induced FKBP and FRB dimerization, in which all parameters were simultaneously estimated using one set of fluorescence resonance energy transfer (FRET) experiments...
June 22, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28630422/basic-surface-features-of-nuclear-fkbps-facilitate-chromatin-binding
#20
Andrew Leung, Francy-Pesek Jardim, Neda Savic, Yoan R Monneau, Rodrigo González-Romero, Geoff Gudavicius, Jose M Eirin-Lopez, Till Bartke, Cameron D Mackereth, Juan Ausió, Christopher J Nelson
The nucleoplasmin family of histone chaperones is identified by a pentamer-forming domain and multiple acidic tracts that mediate histone binding and chaperone activity. Within this family, a novel domain organization was recently discovered that consists of an N-terminal nucleoplasmin-like (NPL) domain and a C-terminal FKBP peptidyl-proline isomerase domain. Saccharomyces cerevisiae Fpr4 is one such protein. Here we report that in addition to its known histone prolyl isomerase activities, the Fpr4 FKBP domain binds to nucleosomes and nucleosome arrays in vitro...
June 19, 2017: Scientific Reports
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