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MicroScale Thermophoresis

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https://www.readbyqxmd.com/read/28721412/effect-of-ligand-binding-on-a-protein-with-a-complex-folding-landscape
#1
Hisham Mazal, Haim Aviram, Inbal Riven, Gilad Haran
Ligand binding to a protein can stabilize it significantly against unfolding. The variation of the folding free energy, ΔΔG(0), due to ligand binding can be derived from a simple reaction scheme involving exclusive binding to the native state. One obtains the following expression: , where Kd is the ligand dissociation constant and L is its concentration, R is the universal gas constant and T is the temperature. This expression has been shown to correctly describe experimental results on multiple proteins...
July 19, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28717148/human-ribosomal-p1-p2-heterodimer-represents-an-optimal-docking-site-for-ricin-a-chain-with-a-prominent-role-for-p1-c-terminus
#2
Przemysław Grela, Xiao-Ping Li, Patrycja Horbowicz, Monika Dźwierzyńska, Marek Tchórzewski, Nilgun E Tumer
The eukaryotic P-stalk contains two P1-P2 protein dimers with a conserved C- terminal domain (CTD) critical for the interaction with external factors. To understand the role of the individual CTD of human P1/P2 proteins, we examined the interaction of reconstituted human P-protein complexes and C-terminally truncated forms with ricin A chain (RTA), which binds to the stalk to depurinate the sarcin/ricin loop (SRL). The interaction between P-protein complexes and RTA was examined by surface plasmon resonance, isothermal titration calorimetry, microscale thermophoresis and bio-layer interferometry...
July 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28710278/phosphorylation-of-human-aquaporin-2-aqp2-allosterically-controls-its-interaction-with-the-lysosomal-trafficking-protein-lip5
#3
Jennifer Virginia Roche, Sabeen Survery, Stefan Kreida, Veronika Nesverova, Henry Ampah-Korsah, Maria Gourdon, Peter M T Deen, Susanna Toernroth-Horsefield
The interaction between the renal water channel aquaporin-2 (AQP2) and the lysosomal trafficking regulator-interacting protein LIP5 targets AQP2 to multivesicular bodies and facilitates lysosomal degradation. This interaction is part of a process which controls AQP2 apical membrane abundance in a vasopressin-dependent manner, allowing for urine volume adjustment. Vasopressin regulates phosphorylation at four sites within the AQP2 C-terminus (S256, S261, S264, T269), of which S256 is crucial and sufficient for AQP2 translocation from storage vesicles to the apical membrane...
July 14, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28698641/a-transcription-activator-like-effector-tal7-of-xanthomonas-oryzae-pv-oryzicola-activates-rice-gene-os09g29100-to-suppress-rice-immunity
#4
Lulu Cai, Yanyan Cao, Zhengyin Xu, Wenxiu Ma, Muhammad Zakria, Lifang Zou, Zaiquan Cheng, Gongyou Chen
Xanthomonas oryzae pv. oryzicola (Xoc) and X. oryzae pv. oryzae (Xoo) cause bacterial leaf streak (BLS) and bacterial leaf blight (BLB) in rice, respectively. Unlike Xoo, endogenous avirulence-resistance (avr-R) gene interactions have not been identified in the Xoc-rice pathosystem; however, both pathogens possess transcription activator-like effectors (TALEs) that are known to modulate R or S genes in rice. The transfer of individual tal genes from Xoc RS105 (hypervirulent) into Xoc YNB0-17 (hypovirulent) led to the identification of tal7, which suppressed avrXa7-Xa7 mediated defense in rice containing an Xa7 R gene...
July 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28681565/tubulin-acetylation-a-novel-functional-avenue-for-cdyl-in-sperm
#5
Sweta Parab, Veena Dalvi, Sushma Mylavaram, Abhipriya K, Susan Idicula-Thomas, Shobha Sonawane, Priyanka Parte
Motility in sperm is driven by the flagella, the principal component of which is the axoneme. The microtubules which make up the 9 + 2 axoneme are composed of heterodimers of alpha and beta tubulins and undergo several post-translational modifications. We have earlier reported that HDAC6 functions as tubulin deacetylase in sperm and has a role in sperm movement. While exploring the specific tubulin acetyltransferase (TAT) in sperm we observed the presence of Chromodomain Y-Like (CDYL), on the principal piece of rat spermatozoa which compelled us to explore its function in sperm...
July 6, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28674022/fsip1-binds-her2-directly-to-regulate-breast-cancer-growth-and-invasiveness
#6
Tong Liu, Hao Zhang, Li Sun, Danyu Zhao, Peng Liu, Meisi Yan, Neeha Zaidi, Sudeh Izadmehr, Animesh Gupta, Wahid Abu-Amer, Minna Luo, Jie Yang, Xunyan Ou, Yining Wang, Xuefeng Bai, Yan Wang, Maria I New, Mone Zaidi, Tony Yuen, Caigang Liu
Fibrous sheath interacting protein 1 (FSIP1), a spermatogenesis-related testicular antigen, is expressed in abundance in breast cancers, particularly in those overexpressing human epidermal growth factor receptor 2 (HER2); however, little is known about its role in regulating the growth and metastasis of breast cancer cells. We and others have shown previously that FSIP1 expression in breast cancer correlates positively with HER2-positivity, recurrence, and metastases and negatively with survival. Here, using coimmunoprecipitation and microscale thermophoresis, we find that FSIP1 binds to the intracellular domain of HER2 directly...
July 3, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28640595/detection-and-characterization-of-small-molecule-interactions-with-fibrillar-protein-aggregates-using-microscale-thermophoresis
#7
Emily Fisher, Yanyan Zhao, Robert Richardson, Malgorzata Janik, Alexander K Buell, Franklin I Aigbirhio, Gergely Tóth
Neurodegenerative diseases such as Parkinson's and Alzheimer's disease share the pathological hallmark of fibrillar protein aggregates. The specific detection of these protein aggregates by positron emission tomography (PET) in the patient brain can yield valuable information for diagnosis and disease progression. However, the identification of novel small compounds that bind fibrillar protein aggregates has been a challenge. In this study, microscale thermophoresis (MST) was applied to assess the binding affinity of known small molecule ligands of α-synuclein fibrils, which were also tested in parallel in a thioflavin T fluorescence competition assay for further validation...
July 6, 2017: ACS Chemical Neuroscience
https://www.readbyqxmd.com/read/28616954/surface-charge-and-particle-size-determine-the-metabolic-fate-of-dendritic-polyglycerols
#8
Kritee Pant, Johanna Pufe, Kristof Zarschler, Ralf Bergmann, Jörg Steinbach, Sabine Reimann, Rainer Haag, Jens Pietzsch, Holger Stephan
Dendritic polyglycerols (dPG) are water soluble, polyether-based nanomaterials which hold great potential in diagnostic as well as therapeutic applications. In order to translate them for in vivo applications, a systematic assessment regarding their cell and tissue interactions as well as their metabolic fate in vivo is a crucial step. Herein, we explore the structure-activity relationship of three different sizes (ca. 3, 5, and 10 nm) of neutral dendritic polyglycerol (dPG) and their corresponding negatively charged sulfate analogs (dPGS) on their in vitro and in vivo characteristics...
June 29, 2017: Nanoscale
https://www.readbyqxmd.com/read/28594954/the-monoclonal-s9-6-antibody-exhibits-highly-variable-binding-affinities-towards-different-r-loop-sequences
#9
Fabian König, Thomas Schubert, Gernot Längst
The monoclonal antibody S9.6 is a widely-used tool to purify, analyse and quantify R-loop structures in cells. A previous study using the surface plasmon resonance technology and a single-chain variable fragment (scFv) of S9.6 showed high affinity (0.6 nM) for DNA-RNA and also a high affinity (2.7 nM) for RNA-RNA hybrids. We used the microscale thermophoresis method allowing surface independent interaction studies and electromobility shift assays to evaluate additional RNA-DNA hybrid sequences and to quantify the binding affinities of the S9...
2017: PloS One
https://www.readbyqxmd.com/read/28580714/marine-alkaloid-oroidin-analogues-with-antiviral-potential-a-novel-class-of-synthetic-compounds-targeting-the-cellular-chaperone-hsp90
#10
Katja-Emilia Lillsunde, Tihomir Tomašič, Danijel Kikelj, Päivi Tammela
Marine organisms and their metabolites are a diverse source of scaffolds for potential pharmacological molecular probes and, less frequently, for pharmaceutical lead compounds. In this study, 157 synthetic analogues of marine sponge-derived alkaloids clathrodin and oroidin were screened against replicon models of two RNA viruses, hepatitis C (HCV) and Chikungunya virus (CHIKV) as part of a larger screening project. Four compounds were found to selectively inhibit the HCV replicon (IC50 1.6-4.6 μM). These belong to the 4,5,6,7-tetrahydrobenzo[1,2-d]thiazole class of compounds originally designed to target the ATP-binding site of bacterial DNA gyrase...
June 5, 2017: Chemical Biology & Drug Design
https://www.readbyqxmd.com/read/28522394/interaction-research-on-an-antiviral-molecule-that-targets-the-coat-protein-of-southern-rice-black-streaked-dwarf-virus
#11
Longlu Ran, Yan Ding, Liangzhi Luo, Xiuhai Gan, Xiangyang Li, Yongzhong Chen, Deyu Hu, Baoan Song
Southern rice black-streaked dwarf virus (SRBSDV) coat protein (P10) is the key protein required for viral transmission and host plant infection and is thus a promising target for anti-SRBSDV agent screening. In this study, P10 was obtained from Escherichia coli through cloning, expression, and purification. The antiviral agent Ningnanmycin was selected as control, and a series of antiviral compounds based on the structural scaffold of ferulic acid were analyzed. Size-exclusion chromatography analysis results showed that compound F27 can alter the aggregation of P10 proteins...
May 15, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28499498/aptatope-mapping-of-the-binding-site-of-a-progesterone-aptamer-on-the-steroid-ring-structure
#12
Vasso Skouridou, Thomas Schubert, Abdulaziz S Bashammakh, Mohammad S El-Shahawi, Abdulrahman O Alyoubi, Ciara K O'Sullivan
In this work we report the mapping of the binding site of the only progesterone aptamer published to date, in an approach referred to as aptatope mapping. By linking the binding data obtained from microscale thermophoresis analysis to the structural differences on the ring structure of a range of steroids, we elucidated the moieties involved in aptamer-progesterone binding. This approach can be further exploited for the characterization of aptamer specificity and ultimately facilitate the development of aptamer-based assays depending on the desired specificity...
August 15, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28459633/identification-of-human-ump-cmp-kinase-1-as-doxorubicin-binding-target-using-protein-microarray
#13
Shuxian Chen, Xu Wang, Xianghui Ye, Donghui Ma, Caiwei Chen, Junlong Cai, Yongfeng Fu, Xunjia Cheng, Yun Chen, Xiaohai Gong, Jian Jin
Doxorubicin (DOX) is a leading anthracycline drug with exceptional efficacy; however, little is known about the molecular mechanisms of its side effects, which include heart muscle damage, noncancerous cell death, and drug resistance. A total of 17,950 human proteins expressed in HEK293 cells were screened and yielded 14 hits. Competitive and binding experiments further verified the binding of DOX to UMP/CMP kinase 1 (CMPK1), and microscale thermophoresis showed that DOX binds to CMPK1 with a Kd of 1216 nM...
April 1, 2017: SLAS Discovery
https://www.readbyqxmd.com/read/28449373/biophysical-analysis-of-the-dynamics-of-calmodulin-interactions-with-neurogranin-and-ca-2-calmodulin-dependent-kinase-ii
#14
Christian Seeger, Vladimir O Talibov, U Helena Danielson
Calmodulin (CaM) functions depend on interactions with CaM-binding proteins, regulated by Ca2+. Induced structural changes influence the affinity, kinetics, and specificities of the interactions. The dynamics of CaM interactions with neurogranin (Ng) and the CaM-binding region of Ca2+/calmodulin-dependent kinase II (CaMKII290-309 ) have been studied using biophysical methods. These proteins have opposite Ca2+ dependencies for CaM binding. Surface plasmon resonance biosensor analysis confirmed that Ca2+ and CaM interact very rapidly, and with moderate affinity ( KDSPR=3μM)...
April 27, 2017: Journal of Molecular Recognition: JMR
https://www.readbyqxmd.com/read/28449085/guanine-quadruplex-monoclonal-antibody-1h6-cross-reacts-with-restrained-thymidine-rich-single-stranded-dna
#15
Hinke G Kazemier, Katrin Paeschke, Peter M Lansdorp
Previously we reported the production and characterization of monoclonal antibody 1H6 raised against (T4G4)2 intermolecular guanine quadruplex (G4) DNA structures (Henderson A. et al. (2014) Nucleic Acids Res., 42, 860-869; Hoffmann R.F. et al. (2016) Nucleic Acids Res., 44, 152-163). It was shown that 1H6 strongly stains nuclei and has exquisite specificity for heterochromatin by immuno-electron microscopy. Here we extend our studies of 1H6 reactivity using enzyme-linked immunosorbent assay (ELISA) and microscale thermophoresis (MST)...
June 2, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28427443/benserazide-a-dopadecarboxylase-inhibitor-suppresses-tumor-growth-by-targeting-hexokinase-2
#16
Wei Li, Mengzhu Zheng, Shuangping Wu, Suyu Gao, Mei Yang, Zhimei Li, Qiuxia Min, Weiguang Sun, Lixia Chen, Guangya Xiang, Hua Li
BACKGROUND: Hexokinase (HK) is the rate-limiting enzyme in the first reaction of glycolysis. And Hexokinase 2 (HK2) is most closely related to malignant tumor which expresses at higher level compared with normal cells. HK2 plays a pivotal role in tumor initiation and maintenance, which provides a new target for cancer therapy. METHODS: Structure-based virtual ligand screening was used in hit identification from ZINC Drug Database. Microscale thermophoresis assay was performed to evaluate the binding affinity...
April 20, 2017: Journal of Experimental & Clinical Cancer Research: CR
https://www.readbyqxmd.com/read/28417365/quantifying-cbm-carbohydrate-interactions-using-microscale-thermophoresis
#17
Haiyang Wu, Cédric Y Montanier, Claire Dumon
MicroScale Thermophoresis (MST) is an emerging technology for studying a broad range of biomolecular interactions with high sensitivity. The affinity constant can be obtained for a wide range of molecules within minutes based on reactions in microliters. Here, we describe the application of MST in quantifying two CBM-carbohydrate interactions, a CBM3a toward cellulose nanocrystals and a CBM4 against xylohexaose.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28404919/paeonol-suppresses-solar-ultraviolet-induced-skin-inflammation-by-targeting-t-lak-cell-originated-protein-kinase
#18
Peipei Xue, Yong Wang, Fanfan Zeng, Ruijuan Xiu, Jingwen Chen, Jinguang Guo, Ping Yuan, Lin Liu, Juanjuan Xiao, Hui Lu, Dan Wu, Huaxiong Pan, Mingmin Lu, Feng Zhu, Fei Shi, Qiuhong Duan
Excessive exposure to solar UV (SUV) is related with numerous human skin disorders, such as skin inflammation, photoaging and carcinogenesis. T-LAK cell- originated protein kinase (TOPK), an upstream of p38 mitogen-activated protein kinase (p38) and c-Jun N-terminal kinases (JNKs), plays an important role in SUV -induced skin inflammation, and targeting TOPK has already been a strategy to prevent skin inflammation. In this study, we found that the expression of TOPK, phosphorylation of p38 or JNKs was increased in human solar dermatitis tissues...
April 18, 2017: Oncotarget
https://www.readbyqxmd.com/read/28344555/drug-repurposing-of-the-anthelmintic-niclosamide-to-treat-multidrug-resistant-leukemia
#19
Sami Hamdoun, Philipp Jung, Thomas Efferth
Multidrug resistance, a major problem that leads to failure of anticancer chemotherapy, requires the development of new drugs. Repurposing of established drugs is a promising approach for overcoming this problem. An example of such drugs is niclosamide, a known anthelmintic that is now known to be cytotoxic and cytostatic against cancer cells. In this study, niclosamide showed varying activity against different cancer cell lines. It revealed better activity against hematological cancer cell lines CCRF-CEM, CEM/ADR5000, and RPMI-8226 compared to the solid tumor cell lines MDA-MB-231, A549, and HT-29...
2017: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/28300592/new-peptide-inhibitors-modulate-the-self-assembly-of-islet-amyloid-polypeptide-residues-11-20-in-vitro
#20
Yexuan Mao, Lanlan Yu, Ran Yang, Chuanguo Ma, Lingbo Qu, Peter de B Harrington
The structural transition and misfolding of human islet amyloid polypeptide may cause a common metabolic disease Type 2 diabetes mellitus. Seventeen peptides have been synthesized, possessing different lengths, compositions, and peptide conformation. In this study, the mechanism of these peptides on inhibiting the formation of hIAPP11-20 amyloid fibrils was investigated using a conventional ThT fluorescence assay and microscale thermophoresis. The results showed that short peptides AT, SA, RF, KS, KT and KN, and cyclic peptides cyclic-KS, cyclic-KT and cyclic-KN displayed considerable inhibitory effect on hIAPP11-20 fibril formation and a strong affinity for hIAPP11-20...
June 5, 2017: European Journal of Pharmacology
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