bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting mER and GLP1R for the Treatment of Atherosclerosis and Type II Diabetes
Authors: Du Jasper , Issa John Paul , Kumar Krishna 
Published: November 2023

Cardiovascular disease (CVD) and type II diabetes (T2D) currently affect hundreds of millions lives worldwide and are at the forefront of public health concerns. The two diseases are interrelated: Type II diabetic patients are at a significantly higher risk of CVD and ultimately succumb to CVD related complications. Estradiol (E2) provides vascular benefits, including vasodilation, reducing the risk of cardiovascular disease. E2 mediates these atheroprotective effects by signaling at the estrogen receptor in the cytosol, endoplasmic reticulum and plasma membrane. In addition, E2 has been seen to have a prominent effect on the hypothalamus, increasing insulin sensitivity and lowering blood glucose levels. Insulin insensitivity is a hallmark of T2D, suggesting that E2 can be a viable molecule to treat T2D. However, binding to cytosolic estrogen receptor (ER) eventually leads to upregulation of proliferative genes that can result in breast or uterine cancers.Glucagon‐like peptide 1 (GLP‐1), a gut‐derived peptidic hormone, resolves high blood sugar levels, body weight, and other T2D complications. Interestingly, GLP‐1 has been observed to have cardiovascular protective effects, reducing risk of CVD. However, GLP‐1 is quickly inactivated in minutes by the endogenous protease dipeptidyl peptidase 4 (DPP‐4) in the bloodstream.These observations point to a link between T2D and CVD, and a link between E2 and GLP‐1. By combining the two hormones together, tangible synergistic benefits can be realized and more importantly, respective obstacles can be avoided. I have developed an in‐solution coupling to chemically link E2 to GLP‐1 at the Lys26 residue. This conjugate would prevent the ability of E2 to initiate breast or uterine cancer but still signal at ER and GLP‐1R. Further, I have identified several methods that prevent proper formation of the conjugate, namely through a Wagner‐Meerwein rearrangement of E2. To preclude inactivation of GLP‐1 by DPP‐4, I have covalently attached a trifluroethyl group at the N‐terminal His of GLP‐1 that retains potency. Additionally, I have conjugated to E2, a small molecule with high affinity to the serum protein transthyretin, in order to extend half‐life and prevent entrance of E2 into the cytosol.Support or Funding InformationI am funded by an ASBMB Travel Award that was presented to me at the Northeastern Regional ASBMB Conference.
Pentabromophenol, a brominated flame retardant derivate, suppresses TGF‐β signaling by sequester TGF‐β receptor from cell surface and further degradation through a caveolae‐mediated endocytosis
Authors: Yang Pei‐hua , Chen Chun‐Lin 
Published: November 2023

Polybormodiphenyl ethers (PBDEs) have been widely used as flame retardants in a variety of consumer products, the possible adverse health effects of PBDEs have focused on their potential endocrine‐disrupting effects and developmental neurotoxicity. In this study, we found that the major biological metabolite of PBDEs, pentabromophenol (PBP) suppressed TGF‐β responsiveness by accelerating the turnover rate of TGF‐β receptor. PBP suppressed TGF‐β1‐mediated cell migration, PAI‐1 reporter gene activation and Smad2/3 activation in various type of cells, abolished TGF‐β1‐mediated repression of E‐cadherin expression, as well as induction of vimentin expression along with Snail and Slug upregulation; thus, blocking the TGF‐β1‐mediated epithelial‐to‐mesenchymal transition (EMT) in A549 calls. TGF‐β superfamily is a key player in the regulation of a wide variety of biological processes from development to pathogenesis including cell proliferation, migration, and the process of cancer development and progression. The in vitro results in this study provide a basis for studies of more detailed relationships between PBP and the modulation of TGF‐β signaling. Given their similarities to PBDEs, additional laboratory and mechanistic study should be considered to assess PBDEs as a potential risk factor for tumorigenesis and other TGF‐β‐related diseases.Support or Funding InformationMinistry of Science and Technology in Taiwan
Unraveling White Lupin's Signal Transduction in Response to Phosphorus Deficiency Using iTRAQ Labeling, Phosphopeptide Enrichment, and Tandem Mass Spectrometry
Authors: Amadi Michael , Cole Jason , Li Kathy , Chalkley Robert J. , Burlingame Al , Uhde‐Stone Claudia 
Published: November 2023

Phosphorus (P) availability is one of the most limiting factors for crop production worldwide. To breed or engineer crops that do well in poor soils, researchers are investigating plants that are well adapted to P deficiency, such as the legume white lupin (Lupinus albus L.). While great progress has been made in understanding white lupin's strategies to acquire P, not much is known about the signal transduction pathways that link sensing of P deficiency with the appropriate responses. Phosphorylation of proteins plays a central role in signaling processes. Proteins that change phosphorylation status when exposed to P deficiency are potential members of such signal transduction pathways. However, quantifying the dynamics of protein phosphorylation is challenging, because of their transient nature and low abundance. A sensitive method is needed to identify proteins that become phosphorylated after exposure to P deficiency.To this end, white lupin was grown in hydroponics with sufficient phosphorus (+P). After 3 weeks, half of the lupin plants were transferred into P‐deficient nutrient solution (−P). After 24 hours, total protein was isolated from roots of two +P and two −P24h plants, for a total of two independent biological replications. Proteins were quantified, trypsin‐digested, and subjected to LC‐MS (Liquid chromatography‐mass spectrometry). This successfully identified hundreds of white lupin proteins, indicating good quality of sample preparation. We then used four different iTRAQ (isobaric tags for relative and absolute quantification) tags to differentially label the +P and −P peptides from the two replicates. We are currently in the process of enriching phosphorylated peptides by titanium dioxide chromatography. Next, we plan to perform multiplex UPLC‐MS/MS (Ultra‐Performance Liquid chromatography tandem mass spectrometry) to quantify the iTRAQ‐labeled phosphorylated peptides from +P and −P24h in two biological replicates. Peptides that are not phosphorylated under +P, but become phosphorylated after transfer to −P are likely members of white lupin's signal transduction pathway in response to P deficiency.
ER/K Linked GPCR‐G protein Fusions Systematically Modulate Second Messenger Response in Cells
Authors: Dysthe Matthew , Malik Rabia U , Ritt Michael , Sunahara Roger K , Sivaramakrishnan Sivaraj 
Published: November 2023

FRET or BRET approaches are well established for detecting ligand induced GPCR‐G protein interactions in cells. However, current FRET/BRET assays rely on co‐expression of GPCR and G protein, and hence depend on the stoichiometry and expression levels of the donor and acceptor probes. On the other hand, GPCR‐G protein fusions have been used extensively to understand the selectivity of GPCR signaling pathways. However, the signaling properties of fusion proteins are not consistent across GPCRs. In this study, we describe novel sensors based on the Systematic Protein Affinity Strength Modulation (SPASM) technique. Sensors consist of a GPCR and G protein tethered by an ER/K linker flanked by FRET probes. SPASM sensors are tested for the beta2‐, alpha1‐, and alpha2‐adrenergic receptors, and adenosine type 1 receptor (A1R), tethered to Gαs‐XL, Gαi1, or Gαq subunits. Agonist stimulation of beta2‐AR and alpha2‐AR increases FRET signal comparable to co‐expressed FRET/BRET sensors. SPASM sensors retain signaling through the endogenous G protein milieu. Importantly, ER/K linker length systematically tunes the GPCR‐G protein interaction, with consequent modulation of second messenger signaling for cognate interactions. SPASM GPCR sensors serve the dual purpose of detecting agonist‐induced changes in GPCR‐G protein interactions, and linking these changes to downstream signaling.Support or Funding InformationThe research conducted here was funded by the American Heart Association Scientist Development Grant (13SDG14270009), the NIH (1DP2 CA186752‐01, 1‐R01‐GM‐105646‐01‐A1) to S.S. and by the American Heart Association Pre‐doctoral Fellowship (14PRE18560010) to R.U.M.
The Golgi Arf‐GEFs Gea1 and Gea2 Integrate Signals to Coordinate Vesicle Formation
Authors: Gustafson Margaret Ann , Fromme J. Christopher 
Published: November 2023

A major challenge for eukaryotic cells is regulating the transport of protein cargoes among compartments and to and from the plasma membrane. How does a cell guarantee the efficient trafficking of appropriate cargoes to the only the right place at the right time? We and others have shown that at the Golgi complex, the cell's sorting house, decisions about packaging cargo rely on coordination of multiple signals.In Saccharomyces cerevisiae (and similarly in other eukaryotes including humans), this coordination relies heavily on small GTPases. The GTPase Arf1 acts as a central molecular switch: when switched on (GTP‐bound), it recruits cargoes, adapters, and coat proteins needed to form a vesicle at the Golgi. Three homologous proteins, called guanine nucleotide exchange factors (Arf‐GEFs), are responsible for exchanging GTP for GDP on Arf1 at the Golgi to activate the molecular switch. Sec7 (BIG1/2 in humans) turns on Arf1 to package secretory cargoes at the trans‐Golgi network (TGN), while Gea1 and Gea2 (GBF1 in humans) help Arf1 package retrograde cargoes into COPI vesicles for intra‐Golgi and Golgi‐to‐ER traffic. We have previously shown that Sec7 utilizes autoinhibition, positive feedback, and interactions with multiple small GTPases to regulate its activity and localization. Identifying the corresponding information used by Gea1/2 to make the decision about when and where to activate Arf1 is crucial to understanding how trafficking is regulated between the three Golgi Arf‐GEFs.We have identified three players in the decision‐making process for Gea1/2. First, all C‐terminal domains of each protein are required for regulating recruitment of the GEFs to membranes in vitro and some are required for viability of cells or cellular localization, highlighting the complex nature of coordinated signals. Second, the composition of the membrane surface is important: unlike Sec7, Gea1/2 prefer neutral to negatively‐charged membranes in vitro, paralleling the more neutral early Golgi compartments and negatively‐charged, phosphatidylserine‐rich TGN in vivo. Lastly, the Rab GTPase Ypt1 (Rab1 in humans) aids in recruiting Gea1/2 to membranes in vitro, in agreement with prior findings that overexpression of Ypt1 can suppress temperature sensitive mutations in Gea1/2.Together, these results begin to reveal the careful orchestration of retrograde Golgi membrane trafficking through coincidence detection. By sensing multiple signals for activation and recruitment, the Golgi Arf‐GEFs are able to carefully calibrate their activity and localization to achieve specificity in cargo packing and timing of vesiculation. Our ongoing work focuses on identifying the signals that fine‐tune compartment specificity within the Golgi and understanding the molecular mechanisms of these complementary interactions.Support or Funding InformationThis work was funded by the NIH (T32 GM07273 and R01 GM098621) and by the Harry and Samuel Mann Outstanding Graduate Student Award.
Variants in the Toll‐interacting protein gene (rs5743899) and susceptibility to Plasmodium falciparum malaria infection in West Africa
Authors: Farid Iman , Funwei Roland I , Agyingi Smith , Snyder Tara , Falade Catherine , Ojurongbe Olusola , Thomas Bolaji N 
Published: November 2023

Malaria remains a very significant disease, with high rates of morbidity and mortality, contributing to economic inequities in endemic countries. The parasite is widely distributed with extensive genetic polymorphisms, increasing rates of drug resistance and lack of an effective vaccine. Dysregulated immune response, especially focused on signaling or pathogen recognition factors in immune system pathway is critical to understanding disease severity or pathophysiology. Toll‐interacting protein (TOLLIP) regulates human toll‐like receptor signaling pathways, and its polymorphisms have been associated with infectious disease. To elucidate whether TOLLIP polymorphisms are associated with Plasmodium falciparum infections in West Africa, we examined the genetic frequency of TOLLIP gene polymorphisms (rs5743899 and rs3750920) in 110 patients, with symptomatic, clinical malaria (diagnosis confirmed by microscopy and polymrease chain reaction) and 371 controls (PCR‐negative), recruited from south‐west Nigeria. Genomic DNA samples between groups were analyzed with a polymerase chain reaction and restriction fragment length polymorphism assay. Our results show that the Tollip rs5743899 mutant variant had the lowest frequency compared to the wild type variant, in addition to an extensive disparity between malaria‐infected patients and uninfected controls. There was a significant difference in the genotypic frequency of the rs5743899 mutant variant between malaria‐infected and control groups respectively in West Africa (13.7% versus 20.8%; p=0.09), but not in the rs3750920 polymorphism (57.3% versus 49.3%; p=0.16). This present observation reveals that Tollip gene rs5743899 mutant variant is potentially associated with Plasmodium falciparum malaria infection in West Africa. Interestingly, the most common haplotype among both groups is H2 (40.4% versus 44.1% for malaria‐infected and uninfected controls respectively), combining the mutant variant for both Tollip polymorphisms. Conclusively, this report shows that susceptibility to malaria infection in West Africa and potential contribution to disease severity or preponderance is significantly associated with host genetic variation, with toll‐interacting protein gene a significant member.Support or Funding InformationFunding provided by College of Health Sciences and Technology Faculty Development and Laboratory Support Fund, Rochester Institute of Technology
Construction of a System for the Study of Protein‐Induced Membrane Tubules
Authors: Rivera Alba Katiria González , Forest Katrina T. 
Published: November 2023

Communication between cells and the environment is crucial to life, and correspondingly cells have evolved complex signaling pathways. For example, bacteria have a repertoire of cellular structures that mediate adhesion, motility, virulence, etc. Some of the well‐studied systems are pili, flagella, and multiple secretion systems. Recently, more attention has been paid to membranous structures such as secreted vesicles and membrane appendages. These structures have been shown to play roles in adhesion, cell‐to‐cell communication, and pathogenesis. Membrane appendages appear to be very diverse structures with possible distinct assembly mechanisms. For instance, membrane appendages can appear as tubular structures with a size range of 20–70 ηm in width that extend from the bacterial membrane for very long distances. These membrane appendages are sometime referred as nanotubes and in some instances they have been shown to play a role in metabolic exchange. One study showed that Delftia sp. can form nanopods as the result of the release of outer membrane vesicles that fused together1. Other membrane tubules have been shown to contain protein filaments, such as sheathed flagella and unnatural pili.In our laboratory we attempted to construct a heterologous system for bacterial pilus assembly. However, we found that under special conditions we could trick E. coli to elaborate structures that resembled nanotubes. In our initial studies we expressed different proteins from the Pseudomonas aeruginosa Type‐4 pilus system in E. coli Bl21‐AI, E. coli BL21 (DE3), or E. coli NovaBlue (DE3) and induced protein expression with IPTG and/or arabinose when necessary. Using negative staining and electron microscopy, we observed tubular structures (about 20–40 ηm wide, and longer than 1 μm). Remarkably although formation of these extensions was dependent on the major component of the pilus, PilA, it was independent of pre‐pilin cleavage by pre‐pilin peptidase; we found long membrane tubules in the presence of a non‐cleavable pilin. This system promises a means to investigate in detail how bacteria can build nanotubes and how this process is affected by the presence of transmembrane proteins.Support or Funding InformationNIH 5 T32 GM008349
Effects of Curcumin on Vesicular Stomatitis Virus (VSV) Infection and Dicer‐1 Expression
Authors: Ahmed Jemil , Tan Yuxiao , Ambegaokar Surendra 
Published: November 2023

Vesicular stomatitis virus (VSV), part of Rhabdoviridae, can cause severe illness in many important livestock, such as horses, cattle, and sheep. Curcumin (CUR) is a polyphenolic extract from the spice turmeric, and has a variety of pharmacological benefits including anti‐inflammatory, antioxidant and anti‐viral activities. However, there is limited work on its effects on VSV infection, or a mechanistic description of its anti‐viral properties. The initial goal of this study was to determine if CUR could restrict VSV infection in Vero cells, and if so, to determine the minimal concentration required for restriction. We observed that 10μM of CUR provided robust inhibition of recombinant VSV‐EGFP infection of Vero cells, as measured via plaque assay and fluorescence, with approximately 33% reduced infection at MOI 0.0002 and nearly 90% reduction at MOI 0.00002 after 24 hours. We hypothesized RNAi mechanisms may be effective at restricting VSV infection, and measured Dicer‐1 mRNA via qPCR. We found cells infected with VSV‐EGFP and treated with CUR had higher levels of Dicer‐1 expression than VSV‐EGFP infected cells treated with vehicle (DMSO); Actin was used as reference gene. Preliminary results indicate CUR alone has limited effect on Dicer‐1 expression in the absence of VSV infection. This may indicate that VSV infection itself alters Dicer‐1 expression; CUR restriction of VSV infection may rescue the effect on Dicer‐1 expression by inhibiting VSV replication through an as yet unidentified mechanism. It is know that VSV infection induces oxidative stress to promote apoptosis, and thus the antioxidant properties of curcumin may be the primary effect on VSV restriction, which future studies will investigate.Support or Funding InformationDept. of Botany & Microbiology, Neuroscience Program, and Summer Science Research Program at Ohio Wesleyan Univ.; Ohio Wesleyan Univ. Theory‐to‐Practice Grant.
Metabolic Stress Drives Keratinocyte Defenses Against Staphylococcus aureus Infection
Authors: Wickersham Matthew Adam , Wachtel Sarah , Lung Tania Wong Fok , Jacquet Rudy , Soong Grace , Richardson Anthony , Parker Dane , Prince Alice 
Published: November 2023

OBJECTIVEHuman skin is commonly colonized and infected by Staphylococcus aureus. Under normal conditions skin is relatively hypoxic. Thus, we postulated that the burden of staphylococcal infection would result in competition for glucose and stimulation of HIF1a signaling to initiate keratinocyte metabolic and immune responses to infection.METHODSThe metabolic activity of primary keratinocytes in response S. aureus metabolic mutants was monitored using a SeaHorse analyzer to measure oxygen consumption and extracellular acidification. A transcriptomic analysis of keratinocyte responses to S. aureus USA300 infected keratinocytes was performed and key results were confirmed by qRT‐PCR and immunoblot. The immunological consequences of Hif1a signaling were demonstrated in vitro, as the metabolic and inflammatory activities of primary keratinocytes were studied through qRT arrays, western blots, ELISA assays and confocal imaging. The in vivo consequences of the glycolytic response to infection were confirmed in vivo in a murine model of skin infection.ABSTRACTUsing a combination of metabolic and transcriptomic methodologies, we found that S. aureus infection is sensed as a metabolic stress by keratinocytes. This is sufficient to induce HIF1a signaling, which promotes IL‐1b production and generates components of the aerobic glycolysis pathway to meet the metabolic requirements of infection. Staphylococci capable of glycolysis, including WT and agr mutants readily induced HIF1a. In contrast, pyk glycolytic mutants failed to compete with keratinocytes for their metabolic needs. Although S. aureus activate the Akt/mTOR cascade, inhibition of this pathway did not inhibit the keratinocyte cytokine response to infection. However, 2‐DG mediated suppression of glycolysis blocked keratinocyte production of IL‐1b in vitro and significantly exacerbated the S. aureus cutaneous infection in a murine model.CONCLUSIONSS. aureus is perceived as a metabolic stress by infected keratinocytes. These keratinocytes respond through HIF1a signaling, increasing glycolytic activity and generating IL‐1b. S. aureus glycolytic mutants, independent of toxin production, fail to activate keratinocytes.Support or Funding InformationFUNDING: The results presented in this abstract were supported by the National Institutes of Health under award number RO1AI103854.
Different Lipids in Synaptic Vesicle and Synaptosome Membrane
Authors: Lewis Kenneth T , Maddipati Krishna R. , Naik Akshata R. , Jena Bhanu P. 
Published: November 2023

Supramolecular 15 nm cup‐shaped lipoprotein structures called porosomes present at the presynaptic membrane, mediate neurotransmitter release via the transient fusion of synaptic vesicles at its base via SNAREs. Major lipids associated with the isolated neuronal porosome complex have been determined to be unique in composition. Since lipids are known to greatly influence various membrane protein association and functions including membrane fusion, the composition of lipids in synaptic vesicle and the synaptosome membrane, was hypothesized to be unique and tightly regulated. To test this hypothesis, the lipidome of isolated synaptosome, synaptosome membrane, and synaptic vesicles, was determined using mass spectrometry. Results from the study demonstrate a unique lipid composition of the synaptic vesicle compared to the synaptosome membrane, reflecting on the tight regulation of nerve cells in maintaining the lipid composition of both organelles.Support or Funding InformationWork presented in this article was supported in part by the National Science Foundation grants EB00303, CBET1066661 (BPJ); the WSU Interdisciplinary Biomedical Systems Fellowship and the Thomas C. Rumble University Graduate Fellowship (KTL); and the WSU Interdisciplinary Biomedical Systems Fellowship (ARN).