journal
https://read.qxmd.com/read/36965620/substrate-recognition-and-proton-coupling-by-a-bacterial-member-of-solute-carrier-family-17
#1
JOURNAL ARTICLE
Samir Batarni, Nanda Nayak, Audrey Chang, Fei Li, Surabhi Hareendranath, Lexi Zhou, Hongfei Xu, Robert Stroud, Jacob Eriksen, Robert H Edwards
The solute carrier 17 (SLC17) family transports diverse organic anions using two distinct modes of coupling to a source of energy. Transporters that package glutamate and nucleotide into secretory vesicles for regulated release by exocytosis are driven by membrane potential but subject to allosteric regulation by H+ and Cl- . Other SLC17 members including the lysosomal sialic acid exporter couple the flux of organic anion to cotransport of H+ . To begin to understand how similar proteins can perform such different functions, we have studied E...
March 23, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36965619/the-wnt-pathway-protein-dvl1-targets-somatostatin-receptor-2-for-lysosome-dependent-degradation
#2
JOURNAL ARTICLE
Heather S Carr, Yan Zuo, Jeffrey A Frost
The Somatostatin receptor 2 (Sstr2) is a heterotrimeric G protein coupled receptor that is highly expressed in neuroendocrine tumors and is a common pharmacological target for intervention. Unfortunately, not all neuroendocrine tumors express Sstr2, and Sstr2 expression can be downregulated with prolonged agonist use. Sstr2 is rapidly internalized following agonist stimulation and, in the short term, is quantitatively recycled back to the plasma membrane. However, mechanisms controlling steady state expression of Sstr2 in the absence of agonist are less well described...
March 23, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36965618/the-batten-disease-protein-cln3-is-important-for-stress-granules-dynamics-and-translational-activity
#3
JOURNAL ARTICLE
Emily L Relton, Nicolas J Roth, Seda Yasa, Abuzar Kaleem, Guido Hermey, Christopher J Minnis, Sara E Mole, Tatyana Shelkovnikova, Stephane Lefrancois, Peter J McCormick, Nicolas Locker
The assembly of membrane-less organelles such as stress granules (SGs) is emerging as central in helping cells rapidly respond and adapt to stress. Following stress sensing, the resulting global translational shutoff leads to the condensation of stalled mRNAs and proteins into SGs. By reorganising cytoplasmic contents, SGs can modulate RNA translation, biochemical reactions and signalling cascades to promote survival until the stress is resolved. While mechanisms for SG disassembly are not widely understood, the resolution of SGs is important for maintaining cell viability and protein homeostasis...
March 23, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36965617/an-affinity-tool-for-the-isolation-of-endogenous-active-mtorc1-from-various-cellular-sources
#4
JOURNAL ARTICLE
Yasir H Ibrahim, Spyridon Pantelios, Anders P Mutvei
The mechanistic target of rapamycin complex 1 (mTORC1) is a central regulator of mammalian cell growth that is dysregulated in a number of human diseases, including metabolic syndromes, aging and cancer. Structural, biochemical and pharmacological studies that have increased our understanding of how mTORC1 executes growth control often relied upon purified mTORC1 protein. However, current immunoaffinity-based purification methods are expensive, inefficient, and do not necessarily isolate endogenous mTORC1, hampering their overall utility in research...
March 23, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36965616/regulatory-mechanism-of-formaldehyde-release-in-heme-degradation-catalyzed-by-staphylococcus-aureus-isdg
#5
JOURNAL ARTICLE
Toshitaka Matsui
IsdG-type enzymes catalyze the non-canonical degradation of heme to iron, staphylobilin (SB), and formaldehyde (HCHO), presumably by binding heme in an unusually distorted conformation. Their unique mechanism has been elucidated for MhuD from Mycobacterium tuberculosis, revealing an unusual ring-opening of hydroxyheme by dioxygenation. A similar mechanism has been postulated for other IsdG enzymes; however, MhuD, which is special as an IsdG-type enzyme, retains a formyl group in the linearized tetrapyrrole...
March 23, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36965615/the-ycx1-protein-encoded-by-the-yeast-ydl206w-gene-plays-a-role-in-calcium-and-calcineurin-signaling
#6
JOURNAL ARTICLE
Grace M Lee, Fangli Weng, Juliana Cranley, Abhinav Rajasekhar, Matthew Stoeckel, Thomas Kane, Renata Tisi, Yuqi Wang
Calcium is ubiquitously present in all living cells and plays important regulatory roles in a wide variety of biological processes. In yeast, many effects of calcium are mediated via the action of calcineurin, a calcium/calmodulin-dependent protein phosphatase. Proper signaling of calcium and calcineurin is important in yeast, and the calcineurin pathway has emerged as a valuable target for developing novel anti-fungal drugs. Here we report a role of YDL206W in calcium and calcineurin signaling in yeast. YDL206W is an uncharacterized gene in yeast, encoding a protein with two sodium/calcium exchange domains...
March 23, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36965614/high-sensitivity-and-low-cost-flavin-luciferase-flux-vc-based-reporter-gene-for-mammalian-cell-expression
#7
JOURNAL ARTICLE
Jittima Phonbuppha, Ruchanok Tinikul, Yoshihiro Ohmiya, Pimchai Chaiyen
Luciferase-based gene reporters generating bioluminescence signals are important tools for biomedical research. Amongst the luciferases, flavin-dependent enzymes use the most economical chemicals. However, their applications in mammalian cells are limited due to their low signals compared to other systems. Here, we constructed Flavin Luciferase from Vibrio campbellii (Vc) for Mammalian Cell Expression (FLUXVc ) by engineering luciferase from Vibrio campbellii (the most thermostable bacterial luciferase reported to date) and optimizing its expression and reporter assays in mammalian cells which can improve the bioluminescence light output by >400-fold as compared to the non-engineered version...
March 23, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963497/identification-of-gpi-anchored-protein-lypd1-as-an-essential-factor-for-odontoblast-differentiation-in-tooth-development
#8
JOURNAL ARTICLE
Yao Fu, Kanako Miyazaki, Yuta Chiba, Keita Funada, Tomomi Yuta, Tian Tian, Kanji Mizuta, Jumpei Kawahara, Ling Zhang, Daniel Martin, Tsutomu Iwamoto, Ichiro Takahashi, Satoshi Fukumoto, Keigo Yoshizaki
Lipid rafts are membrane microdomains rich in cholesterol, sphingolipids, glycosylphosphatidylinositol-anchored proteins (GPI-APs), and receptors. These lipid raft components are localized at the plasma membrane and are essential for signal transmission and organogenesis. However, few reports have been published on the specific effects of lipid rafts on tooth development. Using microarray and single-cell RNA sequencing methods, we found that a GPI-AP, lymphocyte antigen-6 (Ly6)/Plaur domain-containing 1 (Lypd1), was specifically expressed in preodontoblasts...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963496/a-glucose-sensing-mechanism-with-glucose-transporter-1-and-pyruvate-kinase-in-the-area-postrema-regulates-hepatic-glucose-production-in-rats
#9
JOURNAL ARTICLE
Rosa J W Li, Jennifer F M Chiu, Kyla Bruce, Song-Yang Zhang, Daniel R Barros, Jessica T Y Yue, Tony K T Lam
The area postrema (AP) of the brain is exposed to circulating metabolites and hormones. However, whether AP detects glucose changes to exert biological responses remains unknown. Its neighboring nuclei, the nucleus tractus solitarius (NTS), responds to acute glucose infusion by inhibiting hepatic glucose production, but the mechanism also remains elusive. Herein, we characterized AP and NTS glucose-sensing mechanisms. Infusion of glucose into the AP, like the NTS, of chow rats suppressed glucose production during the pancreatic (basal insulin)-euglycemic clamps...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963495/mutagenesis-and-structural-analysis-reveal-the-ctx-m-%C3%AE-lactamase-active-site-is-optimized-for-cephalosporin-catalysis-and-drug-resistance
#10
JOURNAL ARTICLE
Shuo Lu, Miranda Montoya, Liya Hu, Neetu Neetu, Banumathi Sankaran, B V Venkataram Prasad, Timothy Palzkill
CTX-M β-lactamases are a widespread source of resistance to β-lactam antibiotics in Gram-negative bacteria. These enzymes readily hydrolyze penicillins and cephalosporins, including oxyimino-cephalosporins such as cefotaxime. To investigate the preference of CTX-M enzymes for cephalosporins, we examined eleven active-site residues in the CTX-M-14 β-lactamase model system by alanine mutagenesis to assess the contribution of the residues to catalysis and specificity for the hydrolysis of the penicillin, ampicillin, and the cephalosporins cephalothin and cefotaxime...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963494/distinct-effects-of-two-hearing-loss-associated-mutations-in-the-sarcomeric-myosin-myh7b
#11
JOURNAL ARTICLE
Lindsey A Lee, Samantha K Barrick, Ada E Buvoli, Jonathan Walklate, W Tom Stump, Michael Geeves, Michael J Greenberg, Leslie A Leinwand
For decades, sarcomeric myosin heavy chain proteins were assumed to be restricted to striated muscle where they function as molecular motors that contract muscle. However, MYH7b, an evolutionarily ancient member of this myosin family, has been detected in mammalian nonmuscle tissues, and mutations in MYH7b are linked to hereditary hearing loss in compound heterozygous patients. These mutations are the first associated with hearing loss rather than a muscle pathology, and because there are no homologous mutations in other myosin isoforms, their functional effects were unknown...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963493/the-bacterial-nucleoid-associated-proteins-hu-and-dps-condense-dna-into-context-dependent-biphasic-or-multiphasic-complex-coacervates
#12
JOURNAL ARTICLE
Archit Gupta, Ashish Joshi, Kanika Arora, Samrat Mukhopadhyay, Purnananda Guptasarma
The bacterial chromosome, known as its nucleoid, is an amorphous assemblage of globular nucleoprotein domains. It exists in a state of phase separation from the cell's cytoplasm, as an irregularly-shaped, membrane-less, intracellular compartment. This state (the nature of which remains largely unknown) is maintained through bacterial generations ad infinitum. Here, we show that HU, and Dps, two of the most abundant nucleoid-associated proteins (NAPs) of Escherichia coli, undergo spontaneous complex coacervation with different forms of DNA/RNA, both individually and in each other's presence, to cause accretion and compaction of DNA/RNA into liquid-liquid phase separated (LLPS) condensates in vitro...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963492/structure-and-raf-family-kinase-isoform-selectivity-of-type-ii-raf-inhibitors-tovorafenib-and-naporafenib
#13
JOURNAL ARTICLE
Emre Tkacik, Kunhua Li, Gonzalo Gonzalez-Del Pino, Byung Hak Ha, Javier Vinals, Eunyoung Park, Tyler S Beyett, Michael J Eck
Upon activation by RAS, RAF-family kinases initiate signaling through the MAP kinase cascade to control cell growth, proliferation, and differentiation. Among RAF isoforms (ARAF, BRAF, and CRAF), oncogenic mutations are by far most frequent in BRAF. The BRAFV600E mutation drives more than half of all malignant melanoma and is also found in many other cancers. Selective inhibitors of BRAFV600E (vemurafenib, dabrafenib, encorafenib) are used clinically for these indications, but they are not effective inhibitors in the context of oncogenic RAS, which drives dimerization and activation of RAF, nor for malignancies driven by aberrantly dimerized truncation/fusion variants of BRAF...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963491/the-yeast-gdt1-protein-mediates-the-exchange-of-h-for-ca-2-and-mn-2-influencing-the-golgi-ph
#14
JOURNAL ARTICLE
Antoine Deschamps, Louise Thines, Anne-Sophie Colinet, Jiri Stribny, Pierre Morsomme
The GDT1 family is broadly spread and highly conserved among living organisms. GDT1 members have functions in key processes like glycosylation in humans and yeasts, and photosynthesis in plants. These functions are mediated by their ability to transport ions. While transport of Ca2+ or Mn2+ is well established for several GDT1 members, their transport mechanism is poorly understood. Here, we demonstrate that H+ ions are transported in exchange for Ca2+ and Mn2+ cations by the Golgi-localized yeast Gdt1 protein...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963490/inhibition-of-nucleotide-biosynthesis-disrupts-lipid-accumulation-and-adipogenesis
#15
JOURNAL ARTICLE
Abhijit B Shinde, Elizabeth R Nunn, Genesis A Wilson, Mathew T Chvasta, Julia A Pinette, Jacob W Myers, Sun H Peck, Jessica B Spinelli, Elma Zaganjor
Energy balance and nutrient availability are key determinants of cellular decisions to remain quiescent, proliferate or differentiate into a mature cell. After assessing its environmental state, the cell must rewire its metabolism to support distinct cellular outcomes. Mechanistically, how metabolites regulate cell fate decisions is poorly understood. We used adipogenesis as our model system to ascertain the role of metabolism in differentiation. We isolated adipose tissue stromal vascular fraction (SVF) cells and profiled metabolites before and after adipogenic differentiation to identify metabolic signatures associated with these distinct cellular states...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963489/visualizing-the-coordination-of-apurinic-apyrimidinic-endonuclease-ape1-and-dna-polymerase-%C3%AE-during-base-excision-repair
#16
JOURNAL ARTICLE
Max S Fairlamb, Maria Spies, Todd M Washington, Bret D Freudenthal
Base Excision Repair (BER) is carried out by a series of proteins that function in a step-by-step process to identify, remove, and replace DNA damage. During BER, the DNA transitions through various intermediate states as it is processed by each DNA repair enzyme. Left unrepaired, these BER intermediates can transition into double-stranded DNA breaks and promote genome instability. Previous studies have proposed a short-lived complex consisting of the BER intermediate, the incoming enzyme, and the outgoing enzyme at each step of the BER pathway to protect the BER intermediate...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36963488/exploration-of-o-glcnac-transferase-ogt-glycosylation-sites-reveals-a-target-sequence-compositional-bias
#17
JOURNAL ARTICLE
P Andrew Chong, Michael Nosella, Manasvi Vanama, Roxana Ruiz-Arduengo, Julie D Forman-Kay
O-GlcNAc transferase (OGT) is an essential glycosylating enzyme that catalyzes the addition of N-acetylglucosamine to serine or threonine residues of nuclear and cytoplasmic proteins. The enzyme glycosylates a broad range of peptide sequences and prediction of glycosylation sites has proven challenging. The lack of an experimentally verified set of polypeptide sequences that are not glycosylated by OGT has made prediction of legitimate glycosylation sites more difficult. Here, we tested a number of intrinsically disordered protein regions as substrates of OGT to establish a set of sequences that are not glycosylated by OGT...
March 22, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36958475/the-intramembrane-cooh-terminal-domain-of-prrt2-regulates-voltage-dependent-na-channels
#18
JOURNAL ARTICLE
Francesca Franchi, Antonella Marte, Beatrice Corradi, Bruno Sterlini, Giulio Alberini, Alessandra Romei, Antonio De Fusco, Alexander Vogel, Luca Maragliano, Pietro Baldelli, Anna Corradi, Pierluigi Valente, Fabio Benfenati
Proline-rich transmembrane protein 2 (PRRT2) is the single causative gene for pleiotropic paroxysmal syndromes including epilepsy, kinesigenic dyskinesia, episodic ataxia and migraine. PRRT2 is a neuron-specific type-2 membrane protein with a COOH-terminal intramembrane domain and a long proline-rich NH2 -terminal cytoplasmic region. A large array of experimental data indicates that PRRT2 is a neuron stability gene that negatively controls intrinsic excitability by regulating surface membrane localization and biophysical properties of voltage-dependent Na+ channels Nav1...
March 21, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36944399/structural-insights-into-a-cooperative-switch-between-one-and-two-fimh-bacterial-adhesins-binding-pauci-and-high-mannose-type-n-glycan-receptors
#19
JOURNAL ARTICLE
Eva-Maria Krammer, Clarisse Bridot, Sonia Serna, Begoña Echeverria, Shubham Semwal, Benoît Roubinet, Kim van Noort, RuudH P Wilbers, Gleb Bourenkov, Jérôme de Ruyck, Ludovic Landemarre, Niels Reichardt, Julie Bouckaert
The FimH type-1 fimbrial adhesin allows pathogenic Escherichia coli to adhere to glycoproteins in the epithelial linings of human bladder and intestinal tract, by using multiple fimbriae simultaneously. Pauci- and high-mannose type N-glycans are natural FimH receptors on those glycoproteins. Oligomannose-3 and -5 bind with the highest affinity to FimH by using the same Manα1,3Man branch. Oligomannose-6 is generated from oligomannose-5 in the next step of the biogenesis of high-mannose N-glycans, by the transfer of a mannose in α1,2-linkage onto this branch...
March 19, 2023: Journal of Biological Chemistry
https://read.qxmd.com/read/36944398/different-transmembrane-domains-determine-the-specificity-and-efficiency-of-the-cleavage-activity-of-the-%C3%AE-secretase-subunit-presenilin
#20
JOURNAL ARTICLE
FabianC Schmidt, Katja Fitz, LukasP Feilen, Masayasu Okochi, Harald Steiner, Dieter Langosch
The γ-secretase complex catalyzes the intramembrane cleavage of C99, a carboxy-terminal fragment of the amyloid precursor protein (APP). Two paralogs of its catalytic subunit presenilin (PS1 and PS2) are expressed which are autocatalytically cleaved into an N-terminal and a C-terminal fragment (NTF, CTF) during maturation of γ-secretase. In this study, we compared the efficiency and specificity of C99 cleavage by PS1- and PS2-containing γ-secretases. Mass spectrometric analysis of cleavage products obtained in cell-free and cell-based assays revealed that the previously described lower Aβ38 generation by PS2 is accompanied by a reciprocal increase in Aβ37 production...
March 19, 2023: Journal of Biological Chemistry
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