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Jens Heller, Corinne Clavé, Pierre Gladieux, Sven J Saupe, N Louise Glass
In plants and metazoans, intracellular receptors that belong to the NOD-like receptor (NLR) family are major contributors to innate immunity. Filamentous fungal genomes contain large repertoires of genes encoding for proteins with similar architecture to plant and animal NLRs with mostly unknown function. Here, we identify and molecularly characterize patatin-like phospholipase-1 (PLP-1), an NLR-like protein containing an N-terminal patatin-like phospholipase domain, a nucleotide-binding domain (NBD), and a C-terminal tetratricopeptide repeat (TPR) domain...
February 20, 2018: Proceedings of the National Academy of Sciences of the United States of America
Islam M Mostafa, Mahmoud A Omar, Dalia M Nagy, Sayed M Derayea
A simple and sensitive spectrofluorimetric method has been development for the assurance of quetiapine fumarate (QTF). The proposed method was utilized for measuring the fluorescence intensity of the yellow fluorescent product at 510nm (λ ex 470nm). The fluorescent product has resulted from the nucleophilic substitution reaction of QTF with 4-chloro-7-nitrobenzofurazane (NBD-Cl) in Mcllvaine buffer (pH7.0). The diverse variables influencing the development of the reaction product were deliberately changed and optimized...
February 6, 2018: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
Amer Alam, Raphael Küng, Julia Kowal, Robert A McLeod, Nina Tremp, Eugenia V Broude, Igor B Roninson, Henning Stahlberg, Kaspar P Locher
The multidrug transporter ABCB1 (P-glycoprotein) is an ATP-binding cassette transporter that has a key role in protecting tissues from toxic insult and contributes to multidrug extrusion from cancer cells. Here, we report the near-atomic resolution cryo-EM structure of nucleotide-free ABCB1 trapped by an engineered disulfide cross-link between the nucleotide-binding domains (NBDs) and bound to the antigen-binding fragment of the human-specific inhibitory antibody UIC2 and to the third-generation ABCB1 inhibitor zosuquidar...
February 13, 2018: Proceedings of the National Academy of Sciences of the United States of America
Yi Zhong, Jian Feng, Zhongcai Fan, Jiafu Li
Curcumin, which is an extract from a traditional Chinese medicine, has previously been demonstrated to exhibit an anti‑atherosclerotic effect, which is closely associated with an increase in cholesterol efflux. However, it is unclear as to whether the increased effect is mediated by heme oxygenase (HO)‑1. Macrophages were treated with different concentrations of curcumin, HO‑1 inhibitor and small interfering (si)RNA in different experiments. Analysis of protein expression was conducted via western blotting...
February 8, 2018: Molecular Medicine Reports
Zhengrong Yang, Ellen Hildebrandt, Fan Jiang, Andrei A Aleksandrov, Netaly Khazanov, Qingxian Zhou, Jianli An, Andrew T Mezzell, Bala M Xavier, Haitao Ding, John R Riordan, Hanoch Senderowitz, John C Kappes, Christie G Brouillette, Ina L Urbatsch
The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is an ABC transporter containing two transmembrane domains forming a chloride ion channel, and two nucleotide binding domains (NBD1 and NBD2). CFTR has presented a formidable challenge to obtain monodisperse, biophysically stable protein. Here we report a comprehensive study comparing effects of single and multiple NBD1 mutations on stability of both the NBD1 domain alone and on purified full length human CFTR. Single mutations S492P, A534P, I539T acted additively, and when combined with M470V, S495P, and R555K cumulatively yielded an NBD1 with highly improved structural stability...
February 6, 2018: Biochimica et Biophysica Acta
Lance L Goetz, Anton Emmanuel, Klaus Krogh
Introduction: Neurogenic bowel dysfunction (NBD) is a highly prevalent problem after spinal cord injury, with potential for significant impact on health and quality of life. The international standards to document remaining autonomic function after SCI were developed to standardize communication between professionals regarding neurogenic bowel and other autonomic function after SCI. To improve understanding of the bowel subsection, illustrative cases are presented. Case presentation: Three cases are presented which illustrate differences in presentation and scoring of the elements in the data set based upon varying injury severity and location...
2018: Spinal Cord Series and Cases
Fumihiko Igarashi, Mari H Ogihara, Masatoshi Iga, Hiroshi Kataoka
Dietary sterols including cholesterol and phytosterols are essential substrates for insect steroid hormone (ecdysteroid) synthesis in the prothoracic glands (PGs). In a silkworm Bombyx mori, one of the model species of insects, the steroidogenesis has been well demonstrated that cholesterol biotransformation into ecdysone in the PG cells. Because insects lack the ability to synthesize cellular sterol de novo, lipoprotein, lipophorin (Lp), has been thought to be the major cholesterol supply source; however, details of cholesterol behavior from Lp to the PG cells has not been analyzed till date...
January 31, 2018: Steroids
Kristina V Tugaeva, Yaroslav V Faletrov, Elvin S Allakhverdiev, Vladimir M Shkumatov, Eugene G Maksimov, Nikolai N Sluchanko
Steroidogenic acute regulatory protein (StAR, STARD1) is a key factor of intracellular cholesterol transfer to mitochondria, necessary for adrenal and gonadal steroidogenesis, and is an archetypal member of the START protein family. Despite the common overall structural fold, START members differ in their binding selectivity toward various lipid ligands, but the lack of direct structural information hinders complete understanding of the binding process and cholesterol orientation in the STARD1 complex in particular...
February 3, 2018: Biochemical and Biophysical Research Communications
Dániel Szöllősi, Gergely Szakács, Peter Chiba, Thomas Stockner
P-glycoprotein, also known as multidrug resistance protein 1 or ABCB1, can export a wide range of chemically unrelated compounds, including chemotherapeutic drugs. ABCB1 consists of two transmembrane domains that form the substrate binding and translocation domain, and of two cytoplasmic nucleotide binding domains (NBDs) that energize substrate transport by ATP binding and hydrolysis. ATP binding triggers dimerization of the NBDs, which switches the transporter from an inward facing to an outward facing transmembrane domain conformation...
January 23, 2018: Biophysical Journal
Changcheng You, Jianing Zu, Xiaoqi Liu, Pengyu Kong, Chengchao Song, Rongzhi Wei, Changlong Zhou, Yufu Wang, Jinglong Yan
Synovial fibroblasts (SFs) play a crucial role in the inflammatory process of rheumatoid arthritis (RA). The highly activated NF-κB signal in SFs is responsible for most of the synovial inflammation associated with this disease. In this study, we have developed an SF-targeting liposomal system that encapsulates the NF-κB-blocking peptide (NBD peptide) HAP-lipo/NBD. HAP-lipo/NBDs demonstrated efficient SF-specific targeting in vitro and in vivo. Our study also showed a significant inhibitory effect of HAP-lipo/NBD on NF-κB activation, inflammatory cytokine release and SF migration capability after zymosan stimulation...
January 30, 2018: Journal of Cellular and Molecular Medicine
Huan-Huan Sha, Zhen Wang, Shu-Chen Dong, Tian-Mu Hu, Si-Wen Liu, Jun-Ying Zhang, Yang Wu, Rong Ma, Jian-Zhong Wu, Dan Chen, Ji-Feng Feng
The nitrobenzoxadiazole (NBD) derivatives are a series of compounds containing the NBD scaffold that are not glutathione (GSH) peptidomimetics, and result in a strong inhibition of glutathione S-transferases (GSTs). Growing evidences highlight their pivotal roles and outstanding anticancer activity in different tumor models. In particular, 6- (7-nitro-2,1,3-benzoxadiazol-4-ylthio) hexanol (NBDHEX) is extensively studied, which is a very efficient inhibitor of GSTP1-1. It triggers apoptosis in several tumor cell lines and this cytotoxic activity is observed at micro and submicromolar concentrations...
January 22, 2018: Bioscience Reports
Kiera R Flynn, Alessandra Sutti, Lisandra L Martin, M Leigh Ackland, Angel A J Torriero
The understanding of lipid bilayer structure and function has been advanced by the application of molecular fluorophores. However, the effects of these probe molecules on the physicochemical properties of membranes being studied are poorly understood. A quartz crystal microbalance with dissipation monitoring instrument was used in this work to investigate the impact of two commonly used fluorescent probes, 1-palmitoyl-2-{12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl}-sn-glycero-3-phosphocholine (NBD-PC) and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine-B-sulfonyl) (Lis-Rhod PE), on the formation and physicochemical properties of a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine supported lipid bilayer (POPC-SLB)...
January 12, 2018: Biochimica et Biophysica Acta
Santosh Kumar Palanirajan, Ulaganathan Sivagnanam, Sowmiya Murugan, Sathyanarayana N Gummadi
Human phospholipid scramblase 3 (hPLSCR3) is a single pass transmembrane protein that plays a vital role in fat metabolism, mitochondrial function, structure, maintenance and apoptosis. The mechanism of action of scramblases remains still unknown and the role of scramblases in phospholipid translocation is heavily debated. hPLSCR3 is the only member of scramblase family localized to mitochondria and is involved in cardiolipin translocation at the mitochondrial membrane. Direct biochemical evidence of phospholipid translocation by hPLSCR3 is yet to be reported...
June 27, 2017: Biological Chemistry
Roman Kityk, Jürgen Kopp, Matthias P Mayer
Efficient targeting of Hsp70 chaperones to substrate proteins depends on J-domain cochaperones, which in synergism with substrates trigger ATP hydrolysis in Hsp70s and concomitant substrate trapping. We present the crystal structure of the J-domain of Escherichia coli DnaJ in complex with the E. coli Hsp70 DnaK. The J-domain interacts not only with DnaK's nucleotide-binding domain (NBD) but also with its substrate-binding domain (SBD) and packs against the highly conserved interdomain linker. Mutational replacement of contacts between J-domain and SBD strongly reduces the ability of substrates to stimulate ATP hydrolysis in the presence of DnaJ and compromises viability at heat shock temperatures...
January 18, 2018: Molecular Cell
Hiroki Yamada, Kazuyuki Saito, Mitsuhiko Hokari, Shuta Toru
Central nervous system involvement in Behçet's disease (Neuro-Behçet's disease: NBD) has been reported to present diverse clinical and pathological manifestations. A few cases presenting with neurological symptoms preceding other systemic features are difficult to be diagnosed. Here we suggest the clinical benefit of brain biopsy with a case of NBD initially presenting neurological symptoms.
September 2017: ENeurologicalSci
Ashley M Groshong, Abhishek Dey, Irina Bezsonova, Melissa J Caimano, Justin D Radolf
Borrelia burgdorferi is an extreme amino acid (AA) auxotroph whose genome encodes few free AA transporters and an elaborate oligopeptide transport system (B. burgdorferi Opp [BbOpp]). BbOpp consists of five oligopeptide-binding proteins (OBPs), two heterodimeric permeases, and a heterodimeric nucleotide-binding domain (NBD). Homology modeling based on the crystal structure of liganded BbOppA4 revealed that each OBP likely binds a distinct range of peptides. Transcriptional analyses demonstrated that the OBPs are differentially and independently regulated whereas the permeases and NBDs are constitutively expressed...
December 19, 2017: MBio
Kaifan Liang, Lirong Wang, Yuqing Su, Mengyang Liu, Rui Feng, Yanzhi Song, Yihui Deng
The markers are the "revealers" of accelerated blood clearance (ABC) phenomenon. PEGylated nanocarriers labeled with various markers have been used to explore the mechanism of ABC. However, different markers were labeled on different nanocarriers, and the influence of different markers on ABC phenomenon is questionable. In this study, tocopheryl nicotinate (TN), N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (NBD-DPPE), and 1,1'-dioctadecyl-3,3,3',3'-tetra-methylindotricarbocyanine iodide (DiR) were selected as markers...
December 13, 2017: European Journal of Pharmaceutical Sciences
Li-Yang Zhang, Xiao-Lu Ge, Zheng Li, Yong-Jian Tang, Yuan-Yuan Xiong, Xue-Jun Li, Jin-Fang Liu, Si-Yi Wanggou, Chun-Tao Li, Kui Yang, Xin Chen, Zhong-Liang Hu, Yun-Sheng Liu, Zhi-Xiong Liu
Non-functioning pituitary adenomas (NFPAs) are the most frequent pituitary tumors. The elucidation of the mechanisms of aggressive NFPAs in bone destruction is required in order to guide the clinical diagnosis and treatment of NFPAs. In the present study, we investigated the differential proteomics of fibroblasts isolated from clinical specimens of NFPAs with or without bone destruction. Proteomic analysis revealed a group of molecules associated with cytoskeleton organization, including caldesmon, were differentially expressed between fibroblasts isolated from bone destruction NFPAs (BD-NFPAs) and fibroblasts isolated from non-bone destruction NFPAs (NBD-NFPAs)...
December 13, 2017: Scientific Reports
Sui Ouchi, Kazuaki Matsumoto, Maki Okubo, Yuta Yokoyama, Junko Kizu
Colistin sulfate, composed of a mixture of colistin A sulfate (CLA) and colistin B sulfate (CLB), is available for treating life-threatening infections caused by multidrug-resistant Gram-negative bacteria. In this study, the CLA and CLB were quantified separately. Colistin sulfate was extracted from rat plasma with a solid-phase extraction C18 cartridge and reacted with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), and the fluorescent derivatives were subjected to reversed phase high-performance liquid chromatography analysis and used to investigate the pharmacokinetics of CLA and CLB in rat plasma...
December 12, 2017: Biomedical Chromatography: BMC
Naoko Inoue
Reaction conditions for the selective derivatization of three types of aromatic diamines without clean-up in an aqueous solution using 4-(N-chloroformylmethyl-N-methylamino)-7-nitro-2,1,3-benzoxadiazole (NBD-COCl) have been developed for simple, fast, and selective analysis by HPLC. The aromatic amines were derivatized to give di-NBD-CO-aromatic amines at 35°C for 5 min in a buffer solution at pH 5 containing a low acetonitrile concentration (0 - 400 ng/mL, n = 5, r >0.998). The retention times of these di-NBD-CO-aromatic amines were <5 min...
2017: Analytical Sciences: the International Journal of the Japan Society for Analytical Chemistry
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