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Biochemical Journal

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https://www.readbyqxmd.com/read/28320779/the-g2385r-risk-factor-for-parkinson-s-disease-enhances-chip-dependent-intracellular-degradation-of-lrrk2
#1
Iakov N Rudenko, Alice Kaganovich, Rebekah G Langston, Aleksandra Beilina, Kelechi Ndukwe, Ravindran Kumaran, Allissa A Dillman, Ruth Chia, Mark R Cookson
Autosomal dominant mutations in leucine-rich repeat kinase 2 ( LRRK2 ) are associated with Parkinson's disease (PD). Most pathogenic LRRK2 mutations result in amino-acid substitutions in the central ROC-COR-Kinase triple domain and affect enzymatic functions of the protein. However, there are several variants in LRRK2 , including the risk factor G2385R, that impact PD pathogenesis by unknown mechanisms. Previously, we have shown that G2385R LRRK2 has decreased kinase activity in vitro and altered affinity to LRRK2 interactors...
March 20, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28302767/effect-of-different-%C3%AE-subunit-isoforms-on-the-regulation-of-ampk
#2
Robin Willows, Naveenan Navaratnam, Ana Lima, Jon Read, David Carling
AMP-activated protein kinase (AMPK) plays a key role in integrating metabolic pathways in response to energy demand. AMPK activation results in a wide range of downstream responses, many of which are associated with improved metabolic outcome, making AMPK an attractive target for the treatment of metabolic diseases. AMPK is a heterotrimeric complex consisting of a catalytic subunit (α) and two regulatory subunits (β and γ). The γ subunit harbours the nucleotide binding sites and plays an important role in AMPK regulation in response to cellular energy levels...
March 16, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28298475/purification-and-biochemical-characterization-of-npabcg5-nppdr5-a-plant-pleiotropic-drug-resistance-transporter-expressed-in-nicotiana-tabacum-by-2-suspension-cells
#3
Frédéric Toussaint, Baptiste Pierman, Aurélie Bertin, Daniel Lévy, Marc Boutry
Pleiotropic drug resistance (PDR) transporters belong to the ABCG subfamily of ATP binding cassette (ABC) transporters and are involved in the transport of various molecules across plasma membranes. During evolution PDR genes appeared independently in fungi and in plants from a duplication of a half-size ABC gene. The enzymatic properties of purified PDR transporters from yeast have been characterized. This is not the case for any plant PDR transporter, or, incidentally, for any purified plant ABC transporter...
March 15, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28298474/tumor-protein-d52-expression-is-post-transcriptionally-regulated-by-t-cell-intercellular-antigen-tia-1-and-tia-related-protein-via-mrna-stability
#4
Hiromi Motohashi, Yoshiki Mukudai, Chihiro Ito, Kosuke Kato, Toshikazu Shimane, Seiji Kondo, Tatsuo Shirota
Although tumor protein D52 (TPD) family proteins were first identified nearly 20 years ago, their molecular regulatory mechanisms remain unclear. Therefore, we investigated the post-transcriptional regulation of TPD52 family genes. An RNA immunoprecipitation assay showed the potential binding ability of TPD52 family mRNAs to several RNA-binding proteins, and an RNA degradation assay revealed that TPD52 is subject to more prominent post-transcriptional regulation than TPD53 and 54. We subsequently focused on the 3'-untranslated region (3'-UTR) of TPD52 as a cis -acting element in post-transcriptional gene regulation...
March 15, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28280111/usp7-deubiquitinase-controls-hiv-1-production-by-stabilizing-tat-protein
#5
Amjad Ali, Rameez Raja, Sabihur Rahman Farooqui, Shaista Ahmad, Akhil C Banerjea
Deubiquitinases (DUBs) are key regulators of complex cellular processes.  HIV-1 Tat is synthesized early after infection and is mainly responsible for enhancing viral production. Here, we report that one of the DUBs, USP7, stabilized HIV-1 Tat protein through its deubiquitination. Treatment with either general DUB inhibitor (PR-619) or USP7-specific inhibitor (P5091) resulted in Tat protein degradation. USP7-specific inhibitor reduced virus production in latently infected T-lymphocytic cell line J1.1, which produces large amounts of HIV-1 upon stimulation...
March 9, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28280110/tata-complexes-exhibit-a-marked-change-in-organisation-in-response-to-expression-of-the-tatbc-complex
#6
Sarah Marie Smith, Andrew Yarwood, Roland A Fleck, Colin Robinson, Corinne J Smith
The twin arginine translocation (Tat) system is an integral membrane protein complex that accomplishes the remarkable feat of transporting large, fully-folded polypeptides across the inner membrane of bacteria, into the periplasm. In Escherichia coli Tat is comprised of three membrane proteins: TatA, TatB and TatC. How these proteins arrange themselves in the inner membrane to permit passage of Tat substrates, whilst maintaining membrane integrity, is still poorly understood.  TatA is the most abundant component of this complex and facilitates assembly of the transport mechanism...
March 9, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28275114/binding-and-inhibition-of-the-ternary-complex-factor-elk-4-sap1-by-the-adapter-protein-dok-4
#7
Erika Hooker, Cindy Baldwin, Victoria Roodman, Anupam Batra, Tomoko Takano, Serge Lemay
The adapter protein Dok-4 has been reported as both activator and inhibitor of Erk and Elk-1, but lack of knowledge about the identity of its partner molecules has precluded any mechanistic insight into these seemingly conflicting properties. We report that Dok-4 interacts with the transactivation domain of Elk-4  through an atypical PTB domain-mediated interaction. Dok-4 possesses a nuclear export signal and can relocalize Elk-4 from nucleus to cytosol, whereas Elk-4 possesses two nuclear localization signals that restrict interaction with Dok-4...
March 8, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28270545/manganese-induced-turnover-of-tmem165
#8
Sven Potelle, Eudoxie Dulary, Leslie Climer, Sandrine Duvet, Willy Morelle, Dorothée Vicogne, Elodie Lebredonchel, Marine Houdou, Corentin Spriet, Marie-Ange Krzewinski-Recchi, Romain Peanne, André Klein, Geoffroy DE Bettignies, Pierre Morsomme, Gert Matthijs, Thorsten Marquardt, Vladimir Lupashin, Francois Foulquier
TMEM165 deficiencies lead to one of the Congenital Disorders of Glycosylation (CDG), a group of inherited diseases where the glycosylation process is altered. We recently demonstrated that the Golgi glycosylation defect due to TMEM165 deficiency resulted from Golgi manganese homeostasis defect and that Mn2+ supplementation was sufficient to rescue normal glycosylation. In this paper we highlight TMEM165 as a novel Golgi protein sensitive to manganese. When cells were exposed to high Mn2+ concentrations, TMEM165 was degraded in lysosomes...
March 7, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28264989/characterization-of-the-catalytic-properties-of-the-membrane-anchored-metalloprotease-adam9-in-cell-based-assays
#9
Thorsten Maretzky, Steven Swendeman, Elin Mogollon, Gisela Weskamp, Umut Sahin, Karina Reiss, Carl P Blobel
ADAM9 (a disintegrin and metalloprotease9) is a membrane-anchored metalloproteinase that has been implicated in pathological retinal neovascularization and in tumor progression. ADAM9 has constitutive catalytic activity in both biochemical and cell-based assays and can cleave several membrane proteins, including Epidermal-Growth-Factor and Ephrin receptor B4. Yet, little is currently known about the catalytic properties of ADAM9 and its posttranslational regulation and inhibitor profile in cell-based assays...
March 6, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28258151/staphylococcus-aureussdre-captures-the-factor-h-c-terminus-via-a-novel-close-dock-lock-and-latch-mechanism-for-complement-evasion
#10
Yingjie Zhang, Minhao Wu, Tianrong Hang, Chengliang Wang, Ye Yang, Weimin Pan, Jianye Zang, Min Zhang, Xuan Zhang
Complement factor H (CFH) is a soluble complement regulatory protein essential for the downregulation of the alternative pathway upon interaction with specific markers on host cell surface. It recognizes the complement component 3b (C3b) and 3d (C3d) fragments in addition to self cell markers (i.e. glycosaminoglycans (GAG), sialic acid) to distinguish host cells that deserve protection from pathogens that need elimination. The Staphylococcus aureus surface protein serine-aspartate repeat protein E (SdrE) was previously reported to bind human CFH as an immune-evasion tactic...
March 3, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28246335/dynamic-regulation-of-extracellular-atp-in-escherichia-coli
#11
Cora Lilia Alvarez, Gerardo Corradi, Natalia Lauri, Irene Marginedas-Freixa, María Florencia Leal Denis, Nicolás Enrique, Sabina María Mate, Verónica Milesi, Mariano Anibal Ostuni, Vanesa Herlax, Pablo J Schwarzbaum
We studied the kinetics of extracellular ATP (ATPe) in Escherichia coli , and their outer membrane vesicles (OMVs) stimulated with amphipatic peptides melittin (MEL) and mastoparan 7 (MST7). Real time luminometry was used to measure ATPe kinetics, ATP release and ATPase activity. The latter was also determined by following [(32)P]Pi released from [γ-(32)P]ATP. E. coli was studied alone, coincubated with Caco-2 cells or in rat jejunum segments. In E. coli , addition of [γ-(32)P]ATP led to uptake and subsequent hydrolysis of ATPe...
February 28, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28246334/express-incorporation-of-membrane-proteins-from-various-human-cell-types-into-phospholipid-bilayer-nanodiscs
#12
Stefanie Mak, Ruoyu Sun, Michael Schmalenberg, Carsten Peters, Peter B Luppa
Analysis of membrane proteins is still inadequately represented in diagnostics despite their importance as drug targets and biomarkers. One main reason is the difficult handling caused by their insolubility in aqueous buffer solutions. The nanodisc technology was developed to circumvent this challenge and enables analysis of membrane proteins with standard research methods. However, existing nanodisc generation protocols rely on time-consuming membrane isolation and protein purification from overexpression systems...
February 28, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28246333/distinct-electron-transfer-from-ferredoxin-thioredoxin-reductase-to-multiple-thioredoxin-isoforms-in-chloroplasts
#13
Keisuke Yoshida, Toru Hisabori
Thiol-based redox regulation is considered to support light-responsive control of various chloroplast functions. The redox cascade via ferredoxin-thioredoxin reductase (FTR)/thioredoxin (Trx) has been recognized as a key to transmitting reducing power; however, the Arabidopsis thaliana genome sequencing has revealed that as many as five Trx subtypes encoded by a total of ten nuclear genes are targeted to chloroplasts. Because each Trx isoform seems to have a distinct target selectivity, the electron distribution from FTR to multiple Trxs is thought to be the critical branch point for determining the consequence of chloroplast redox regulation...
February 28, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28232500/pak5-mediates-cell-cell-adhesion-integrity-via-interaction-with-e-cadherin-in-bladder-cancer-cells
#14
Ahmad Fahim Ismail, Sevil Oskay, Nouf Babteen, Mario De Piano, Tracey Martin, Wen G Jiang, Muhammad Khan, Prokar Dasgupta, Claire M Wells
Urothelial bladder cancer  is a major cause of morbidity and mortality worldwide, causing an estimated 150,000 deaths per year. Whilst non-muscle-invasive bladder tumours can be effectively treated, with high survival rates,  many tumours recur, and some will progress to muscle-invasive disease with a much poorer long term prognosis. Thus there is a pressing need to understand the molecular transitions occurring within the progression of bladder cancer to an invasive disease. Tumour invasion is often associated with a down regulation of E-cadherin expression concomitant with a suppression of cell: cell junctions and decreased levels of E-cadherin expression have been reported in higher grade urothelial bladder tumours...
February 23, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28219939/peroxiredoxin-1-prx1-is-a-dual-function-enzyme-by-possessing-cys-independent-catalase-like-activity
#15
Cen-Cen Sun, Wei-Ren Dong, Tong Shao, Jiang-Yuan Li, Jing Zhao, Li Nie, Li-Xin Xiang, Guan Zhu, Jian-Zhong Shao
Peroxiredoxin (Prx) was previously known as a Cys-dependent thioredoxin. However, we unexpected observed that Prx1 from the green spotted puffer fish Tetraodon nigroviridis (TnPrx1) was able to reduce H2O2 in a manner independent on the Cys peroxidation and reductants. This study aimed to validate the novel function for Prx1, delineate the biochemical features and explore its antioxidant role in cells. We have confirmed that Prx1 from the puffer fish and humans truly possesses a catalase-like activity that is independent of Cys residues and reductants, but dependent on iron...
February 20, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28202711/structural-interface-between-lrrk2-and-14-3-3-protein
#16
Loes Stevers, Rens de Vries, Richard Doveston, Lech-Gustav Milroy, Luc Brunsveld, Christian Ottmann
Binding of 14-3-3 proteins to LRRK2 is known to be impaired by a number of Parkinson's disease (PD)-relevant mutations. Abrogation of this interaction is connected to enhanced LRRK2 kinase activity which in turn is implicated in increased ubiquitination of LRRK2, accumulation of LRRK2 into inclusion bodies and reduction of neurite length. Hence, the interaction between 14-3-3 and LRRK2 is of significant interest as a possible drug target for the treatment of PD. However, LRRK2 possesses multiple sites that upon phosphorylation can bind to 14-3-3, thus rendering the interaction relatively complex...
February 15, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28196833/characterization-of-the-activation-of-small-gtpases-by-their-gefs-on-membranes-using-artificial-membrane-tethering
#17
François Peurois, Simon Veyron, Yann Ferrandez, Ilham Ladid, Sarah Benabdi, Mahel Zeghouf, Gérald Peyroche, Jacqueline Cherfils
Attachment of active, GTP-bound small GTPases to membranes by post-translational lipid modifications is pivotal for their ability to process and propagate information in cells. However, generating and manipulating lipidated GTPases has remained difficult, which has limited our quantitative understanding of their activation by GEFs and their termination by GAPs. Here we replaced the lipid modification by a histidine tag in eleven full-length, human small GTPases belonging to the Arf, Rho and Rab families, which allowed to tether them to nickel-lipid containing membranes and characterize the kinetics of their activation by GEFs...
February 14, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28188256/mgdg-pg-and-sqdg-regulate-the-activity-of-light-dependent-protochlorophyllide-oxidoreductase
#18
Michal Gabruk, Beata Myśliwa-Kurdziel, Jerzy Kruk
Light-dependent protochlorophyllide oxidoreductase (POR) is a plant enzyme involved in the chlorophyll biosynthesis pathway. POR reduces one of the double bonds of the protochlorophyllide (Pchlide) using NADPH and light. In the present paper, we found out that PG and SQDG are allosteric regulators of the nucleotide binding, which increase the affinity towards NADPH a hundred fold. Moreover, we showed for the first time that NADH can, like NADPH, form active complexes with Pchlide and POR, however at much higher concentrations...
February 10, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28188255/efficiacy-of-safranal-to-cisplatin-induced-nephrotoxicity
#19
Yasemin Karafakıoglu, Mehmet Fatih Bozkurt, Ömer Hazman, Fatih Fidan
The aim of this study was to investigate the effects of safranal on cisplatin-induced nephrotoxicity and oxidative stress in rats. Adult male Sprague-Dawley rats were randomly divided into five groups. The control group received physiological saline; animals in group 2 received only safranal and in group 3 received only cisplatin; 5 days of safranal treatment was performed following administration of cisplatin for the animals in group 4;5 days of safranal pre-treatment was applied to the animals in group 5 before administration of cisplatin...
February 10, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28183985/characterization-of-pph3-mediated-dephosphorylation-of-rad53-during-methyl-methanesulfonate-induced-dna-damage-repair-in-candida-albicans
#20
Guangyin Yao, Junhua Wan, Qizheng Liu, Chunhua Mu, Yue Wang, Jianli Sang
Genotoxic stress causes DNA damage or stalled DNA replication and filamentous growth in the pathogenic fungus Candida albicans The DNA checkpoint kinase Rad53 critically regulates by phosphorylation effectors that execute the stress response. Rad53 itself is activated by phosphorylation and inactivated by dephosphorylation. Previous studies have suggested that the phosphatase Pph3 dephosphorylates Rad53. Here, we used mass spectrometry and mutagenesis to identify Pph3 dephosphorylation sites on Rad53 in C. albicans We found that serine residues 351, 461, and 477, which were dephosphorylated in wild-type cells during the recovery from DNA damage caused by methyl methanesulfonate (MMS), remained phosphorylated in pph3Δ/Δ cells...
February 9, 2017: Biochemical Journal
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