journal
https://read.qxmd.com/read/38032258/the-role-of-phosphorylation-in-calmodulin-mediated-gating-of-human-aqp0
#1
JOURNAL ARTICLE
Stefan Kreida, Jennifer Virginia Roche, Julie Winkel Missel, Tamim Al-Jubair, Carl Johan Hagströmer, Veronika Wittenbecher, Sara Linse, Pontus Gourdon, Susanna Törnroth-Horsefield
Aquaporin-0 (AQP0) is the main water channel in the mammalian lens and is involved in accommodation and maintaining lens transparency. AQP0 binds the Ca2+-sensing protein calmodulin (CaM) and this interaction is believed to gate its water permeability by closing the water-conducting pore. Here we express recombinant and functional human AQP0 in P. pastoris and investigate how phosphorylation affects the interaction with CaM in vitro as well as the CaM-dependent water permeability of AQP0 in proteoliposomes...
November 30, 2023: Biochemical Journal
https://read.qxmd.com/read/38014506/sperm-induce-a-secondary-increase-in-atp-levels-in-mouse-eggs-that-is-independent-of-ca2-oscillations
#2
JOURNAL ARTICLE
Cindy Ikie-Eshalomi, Elnur Aliyev, Sven Hoehn, Thomasz Jukowski, Karl Swann
Egg activation at fertilization in mouse eggs is caused by a series of cytosolic Ca2+ oscillations that are associated with an increase in ATP concentrations driven by increased mitochondrial activity. We have investigated the role of Ca2+ oscillations in these changes in ATP at fertilization by measuring the dynamics of ATP and Ca2+ in mouse eggs. An initial ATP increase started with the first Ca2+ transient at fertilization and then a secondary increase in ATP occurred about 1 hour later and this preceded a small and temporary increase in the frequency of Ca2+ oscillations...
November 28, 2023: Biochemical Journal
https://read.qxmd.com/read/38014500/leukotriene-b4-receptor-1-blt1-does-not-mediate-disease-progression-in-a-mouse-model-of-liver-fibrosis
#3
JOURNAL ARTICLE
Erin Coyne, Yilin Nie, Desiree Abdurrachim, Charlene Lin Zhi Ong, Yongqi Zhou, Asad Abu Bakar Ali, Stacey Meyers, Jeff Grein, Wendy Blumenschein, Brendan Gongol, Yang Liu, Cedric Lorenz Hugelshofer, Ester Carballo-Jane, Saswata Talukdar
MASH is a prevalent liver disease that can progress to fibrosis, cirrhosis, hepatocellular carcinoma (HCC), and ultimately death, but there are no approved therapies. Leukotriene B4 (LTB4) is a potent pro-inflammatory chemoattractant that drives macrophage and neutrophil chemotaxis, and genetic loss or inhibition of its high affinity receptor, leukotriene B4 receptor 1 (BLT1), results in improved insulin sensitivity and decreased hepatic steatosis. To validate the therapeutic efficacy of BLT1 inhibition in an inflammatory and pro-fibrotic mouse model of MASH and fibrosis, mice were challenged with a choline-deficient, L-amino acid defined high fat diet and treated with a BLT1 antagonist at 30 or 90 mg/kg for 8 weeks...
November 28, 2023: Biochemical Journal
https://read.qxmd.com/read/37943248/elucidating-the-interaction-of-c-terminal-domain-of-vaccinia-related-kinase2a-vrk2a-with-b-cell-lymphoma-extra-large-bcl-xl-to-decipher-its-anti-apoptotic-role-in-cancer
#4
JOURNAL ARTICLE
Rashmi Puja, Shubhankar Dutta, Kakoli Bose
Vaccinia-Related Kinase 2 (VRK2) is an anti-apoptotic Ser/Thr kinase that enhances drug sensitivity in cancer cells. This protein exists in two isoforms: VRK2A, the longer variant, and VRK2B, which lacks the C-terminal region and transmembrane domain. While the therapeutic importance of VRK2 family proteins is known, the specific roles of VRK2A and its interplay with apoptotic regulator Bcl-xL (B-cell lymphoma-extra Large) remain elusive. Bcl-xL regulates cell death by interacting with BAX (B-cell lymphoma-2 Associated X-protein), controlling its cellular localization and influencing BAX-associated processes and signalling pathways...
November 9, 2023: Biochemical Journal
https://read.qxmd.com/read/37905668/loss-of-the-methylarginine-reader-function-of-snd1-confers-resistance-to-hepatocellular-carcinoma
#5
JOURNAL ARTICLE
Tanner Wright, Yalong Wang, Sabrina A Stratton, Manu Sebastian, Bin Liu, David G Johnson, Mark T Bedford
Staphylococcal nuclease Tudor domain containing 1 (SND1) protein is an oncogene that "reads" methylarginine marks through its Tudor domain. Specifically, it recognizes methylation marks deposited by protein arginine methyltransferase 5 (PRMT5), which is also known to promote tumorigenesis. Although SND1 can drive hepatocellular carcinoma (HCC), it is unclear whether the SND1 Tudor domain is needed to promote HCC. We sought to identify the biological role of the SND1 Tudor domain in normal and tumorigenic settings by developing two genetically engineered SND1 mouse models, an Snd1 knockout (Snd1 KO) and an Snd1 Tudor domain-mutated (Snd1 KI) mouse, whose mutant SND1 can no longer recognize PRMT5-catalyzed methylarginine marks...
October 31, 2023: Biochemical Journal
https://read.qxmd.com/read/37903000/biochemical-investigations-of-polyphenol-degradation-enzymes-in-the-phototrophic-bacterium-rubrivivax-gelatinosus
#6
JOURNAL ARTICLE
Yan Zhou, Mengyu Cui, Yifeng Wei, Jason Tan, Tong Li, Xinan Jiao
Phloroglucinol (1,3,5-trihydroxybenzene) is an important intermediate in the degradation of flavonoids and tannins by anaerobic bacteria. Recent studies have shed light on the enzymatic mechanism of phloroglucinol degradation in butyrate-forming anaerobic bacteria, including environmental and intestinal bacteria such as Clostridium and Flavonifractor sp. Phloroglucinol degradation gene clusters have also been identified in other metabolically diverse bacteria, although the polyphenol metabolism of these microorganisms remain largely unexplored...
October 30, 2023: Biochemical Journal
https://read.qxmd.com/read/37888776/phf1-compartmentalizes-prc2-via-phase-separation
#7
JOURNAL ARTICLE
Genzhe Lu, Pilong Li
Polycomb repressive complex 2 (PRC2) is central to polycomb repression as it trimethylates lysine 27 on histone H3 (H3K27me3). How PRC2 is recruited to its targets to deposit H3K27me3 remains an open question. Polycomb-like (PCL) proteins, a group of conserved PRC2 accessory proteins, can direct PRC2 to its targets. In this report, we demonstrate that a PCL protein named PHF1 forms phase-separated condensates at H3K27me3 loci that recruit PRC2. Combining cellular observation and biochemical reconstitution, we show that the N-terminal domains of PHF1 cooperatively mediate target recognition, the chromo-like domain recruits PRC2, and the intrinsically disordered region (IDR) drives phase separation...
October 27, 2023: Biochemical Journal
https://read.qxmd.com/read/37870100/catalysis-of-non-canonical-protein-ubiquitylation-by-the-arih1-ubiquitin-ligase
#8
JOURNAL ARTICLE
Nicholas Purser, Ishita Tripathi-Giesgen, Jerry Li, Daniel C Scott, Daniel Horn-Ghetko, Khee Baek, Brenda Schulman, Arno Alpi, Gary Kleiger
Protein ubiquitylation typically involves isopeptide bond formation between the C-terminus of ubiquitin to the side-chain amino group on Lys residues. However, several ubiquitin ligases (E3s) have recently been identified that ubiquitylate proteins on non-Lys residues. For instance, HOIL-1 belongs to the RING-in-between RING (RBR) class of E3s and has an established role in Ser ubiquitylation. Given the homology between HOIL-1 and ARIH1, an RBR E3 that functions with the large superfamily of cullin-RING E3 ligases (CRLs), a biochemical investigation was undertaken, showing ARIH1 catalyzes Ser ubiquitylation to CRL-bound substrates...
October 23, 2023: Biochemical Journal
https://read.qxmd.com/read/37869794/activating-mutations-drive-human-mek1-kinase-using-a-gear-shifting-mechanism
#9
JOURNAL ARTICLE
Keshav Patil, Yiming Wang, Zhangtao Chen, Krishna Suresh, Ravi Radhakrishnan
There is an unmet need to classify cancer-promoting kinase mutations in a mechanistically cognizant way. The challenge is to understand mutations stabilize different kinase configurations to alter function, and how this influences pathogenic potential of the kinase and its responses to therapeutic inhibitors. This goal is made more challenging by the complexity of the mutational landscape of diseases, and is further compounded by the conformational plasticity of each variant where multiple conformations co-exist...
October 23, 2023: Biochemical Journal
https://read.qxmd.com/read/37818922/ube2a-b-is-the-trans-acting-factor-mediating-mechanotransduction-and-contact-inhibition
#10
JOURNAL ARTICLE
Mingwei Feng, Jiale Wang, Kangjing Li, Fumihiko Nakamura
Mechanotransduction and contact inhibition (CI) control gene expression to regulate proliferation, differentiation, and even tumorigenesis of cells. However, their downstream trans-acting factors (TAFs) are not well known due to a lack of a high-throughput method to quantitatively detect them. Here, we developed a method to identify TAFs on the cis-acting sequences that reside in open chromatin or DNase I-hypersensitive sites (DHSs) and to detect nucleocytoplasmic shuttling TAFs using computational and experimental screening...
October 11, 2023: Biochemical Journal
https://read.qxmd.com/read/37815456/microrchidia-2-histone-deacetylase-1-complex-regulates-e-cadherin-gene-expression-and-function
#11
JOURNAL ARTICLE
Liz Thomas, Namita Chutani, Krishna R, Asha S Nair, Nanda Kumar Yellapu, Karyala Prashanthi, Suresh Babu Pakala
Although Microrchidia 2 (MORC2) is widely overexpressed in human malignancies and linked to cancer cell proliferation, metabolism, and metastasis, the mechanism of action of MORC2 in cancer cell migration and invasion is yet undeciphered. Here, we identified for the first time that MORC2, a chromatin remodeler, regulates E-cadherin expression and, subsequently regulates breast cancer cell migration and invasion.  We observed a negative correlation between the expression levels of MORC2 and E-cadherin in breast cancer...
October 10, 2023: Biochemical Journal
https://read.qxmd.com/read/37767715/comprehensive-comparative-assessment-of-the-arabidopsis-thaliana-mlo2-calmodulin2-interaction-by-various-in-vitro-and-in-vivo-protein-protein-interaction-assays
#12
JOURNAL ARTICLE
Kira von Bongartz, Björn Sabelleck, Anežka Baquero Forero, Hannah Kuhn, Franz Leissing, Ralph Panstruga
Mildew resistance locus o (MLO) proteins are heptahelical integral membrane proteins of which some isoforms act as susceptibility factors for the powdery mildew pathogen. In many angiosperm plant species, loss-of-function mlo mutants confer durable broad-spectrum resistance against the fungal disease. Barley Mlo is known to interact via a cytosolic carboxyl-terminal domain with the intracellular calcium sensor calmodulin (CAM) in a calcium-dependent manner. Site-directed mutagenesis has revealed key amino acid residues in the barley Mlo calmodulin-binding domain (CAMBD) that, when mutated, affect the MLO-CAM association...
September 28, 2023: Biochemical Journal
https://read.qxmd.com/read/37756534/isgylation-independent-protection-of-cell-growth-by-usp18-following-interferon-stimulation
#13
JOURNAL ARTICLE
Anne Clancy, Emma V Rusilowicz-Jones, Iona Wallace, Kirby Swatek, Sylvie Urbe, Michael J Clague
Type 1 interferon stimulation highly up-regulates all elements of a ubiquitin-like conjugation system that leads to ISGylation of target proteins. An ISG15-specific member of the deubiquitylase family, USP18, is up-regulated in a co-ordinated manner. USP18 can also provide a negative feedback by inhibiting JAK-STAT signaling through protein interactions independently of DUB activity. Here, we provide an acute example of this phenomenon, whereby the early expression of USP18, post-interferon treatment of HCT116 colon cancer cells is sufficient to fully suppress the expression of the ISG15 E1 enzyme, UBA7...
September 26, 2023: Biochemical Journal
https://read.qxmd.com/read/37747814/proteotoxic-stresses-stimulate-dissociation-of-ubl4a-from-the-tail-anchored-protein-recognition-complex
#14
JOURNAL ARTICLE
Hiroyuki Kawahara, Takumi Hagiwara, Ryosuke Minami, Chizuru Ushio, Naoto Yokota
Inclusion body formation is associated with cytotoxicity in a number of neurodegenerative diseases. However, the molecular basis of the toxicity caused by the accumulation of aggregation-prone proteins remains controversial. In this study, we found that disease-associated inclusions induced by elongated polyglutamine chains disrupt the complex formation of BAG6 with UBL4A, a mammalian homologue of yeast Get5. UBL4A also dissociated from BAG6 in response to proteotoxic stresses such as proteasomal inhibition and mitochondrial depolarization...
September 25, 2023: Biochemical Journal
https://read.qxmd.com/read/37747806/genetic-modulation-of-cwd-prion-propagation-in-cervid-prp-drosophila
#15
JOURNAL ARTICLE
Alana M Thackray, Erin E McNulty, Amy V Nalls, Alzbeta Cardova, Linh Tran, Glenn Telling, Sylvie L Benestad, Sabine Gilch, Candace K Mathiason, Raymond Bujdoso
Chronic wasting disease is a fatal prion condition of cervids such as deer, elk, moose and reindeer. Secretion and excretion of prion infectivity from North American cervids with this condition causes environmental contamination and subsequent efficient lateral transmission in free-ranging and farmed cervids. Variants of cervid PrP exist that affect host susceptibility to chronic wasting disease. Cervid breeding programmes aimed at increasing the frequency of PrP variants associated with resistance to chronic wasting disease may reduce the burden of this condition in animals and lower the risk of zoonotic disease...
September 25, 2023: Biochemical Journal
https://read.qxmd.com/read/37747786/functional-annotation-of-haloacid-dehalogenase-superfamily-structural-genomics-proteins
#16
JOURNAL ARTICLE
Lakindu S Pathira Kankanamge, Lydia A Ruffner, Mong Mary Touch, Manuel Pina, Penny J Beuning, Mary Jo Ondrechen
Haloacid dehalogenases (HAD) are members of a large superfamily that includes many Structural Genomics proteins with poorly characterized functionality. This superfamily consists of multiple types of enzymes that can act as sugar phosphatases, haloacid dehalogenases, phosphonoacetaldehyde hydrolases, ATPases, or phosphate monoesterases. Here we report on predicted functional annotations and experimental testing by direct biochemical assay for Structural Genomics proteins from the HAD superfamily. To characterize the functions of HAD superfamily members, nine representative HAD proteins and 21 structural genomics proteins are analyzed...
September 23, 2023: Biochemical Journal
https://read.qxmd.com/read/37721041/crispr-cas9-based-edition-of-frataxin-gene-in-dictyostelium-discoideum
#17
JOURNAL ARTICLE
Hernan Gustavo Gentili, María Florencia Pignataro, Justo Facundo Olmos, María Florencia Paván, Lorena Itatí Ibáñez, Javier Santos, Francisco Velazquez Duarte
In this paper we describe the development of a Dictyostelium discoideum strain deficient in frataxin protein (FXN). We investigated the conservation of function between humans and D. discoideum and showed that DdFXN can substitute the human version in the interaction and activation of the Fe-S assembly supercomplex. We edited the D. discoideum fxn locus and isolated a defective mutant, clone 8, which presents landmarks of frataxin deficiency, such as a decrease in Fe-S cluster-dependent enzymatic functions, growth rate reduction, and increased sensitivity to oxidative stress...
September 18, 2023: Biochemical Journal
https://read.qxmd.com/read/37702403/lhs1-dependent-erad-is-determined-by%C3%A2-transmembrane-domain-context
#18
JOURNAL ARTICLE
Maria Sukhoplyasova, Abigail M Keith, Emma M Perrault, Hannah E Vorndran, Alexa S Jordahl, Megan E Yates, Ashutosh Pastor, Zachary Li, Michael L Freaney, Riddhi A Deshpande, David B Adams, Christopher Guerriero, Shujie Shi, Thomas R Kleyman, Ossama Kashlan, Jeffrey L Brodsky, Teresa M Buck
Transmembrane proteins have unique requirements to fold and integrate into the endoplasmic reticulum (ER) membrane. Most notably, transmembrane proteins must fold in three separate environments: extracellular domains fold in the oxidizing environment of the ER lumen, transmembrane domains (TMDs) fold within the lipid bilayer, and cytosolic domains fold in the reducing environment of the cytosol. Moreover, each region is acted upon by a unique set of chaperones and monitored by components of the ER associated quality control machinery that identify misfolded domains in each compartment...
September 13, 2023: Biochemical Journal
https://read.qxmd.com/read/38038975/a-guide-to-erk-dynamics-part-2-downstream-decoding
#19
JOURNAL ARTICLE
Abhineet Ram, Devan Murphy, Nicholaus DeCuzzi, Madhura Patankar, Jason Hu, Michael Pargett, John G Albeck
Signaling by the extracellular signal-regulated kinase (ERK) pathway controls many cellular processes, including cell division, death, and differentiation. In this second installment of a two-part review, we address the question of how the ERK pathway exerts distinct and context-specific effects on multiple processes. We discuss how the dynamics of ERK activity induce selective changes in gene expression programs, with insights from both experiments and computational models. With a focus on single-cell biosensor-based studies, we summarize four major functional modes for ERK signaling in tissues: adjusting the size of cell populations, gradient-based patterning, wave propagation of morphological changes, and diversification of cellular gene expression states...
December 13, 2023: Biochemical Journal
https://read.qxmd.com/read/38038974/a-guide-to-erk-dynamics-part-1-mechanisms-and-models
#20
JOURNAL ARTICLE
Abhineet Ram, Devan Murphy, Nicholaus DeCuzzi, Madhura Patankar, Jason Hu, Michael Pargett, John G Albeck
Extracellular signal-regulated kinase (ERK) has long been studied as a key driver of both essential cellular processes and disease. A persistent question has been how this single pathway is able to direct multiple cell behaviors, including growth, proliferation, and death. Modern biosensor studies have revealed that the temporal pattern of ERK activity is highly variable and heterogeneous, and critically, that these dynamic differences modulate cell fate. This two-part review discusses the current understanding of dynamic activity in the ERK pathway, how it regulates cellular decisions, and how these cell fates lead to tissue regulation and pathology...
December 13, 2023: Biochemical Journal
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