scholarly journals The infantile myofibromatosis NOTCH3 L1519P mutation leads to hyperactivated ligand-independent Notch signaling and increased PDGFRB expression

2021 ◽  
Vol 14 (2) ◽  
pp. dmm046300
Author(s):  
Dan Wu ◽  
Sailan Wang ◽  
Daniel V. Oliveira ◽  
Francesca Del Gaudio ◽  
Michael Vanlandewijck ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Extracellular Domain ◽  
Infantile Myofibromatosis ◽  
Independent Manner ◽  
Functional Link ◽  
Downstream Signaling ◽  
Chloroquine Treatment ◽  
Notch3 Mutation ◽  
The Relationship

ABSTRACTInfantile myofibromatosis (IMF) is a benign tumor form characterized by the development of nonmetastatic tumors in skin, bone, muscle and sometimes viscera. Autosomal-dominant forms of IMF are caused by mutations in the PDGFRB gene, but a family carrying a L1519P mutation in the NOTCH3 gene has also recently been identified. In this study, we address the molecular consequences of the NOTCH3L1519P mutation and the relationship between Notch and PDGFRB signaling in IMF. The NOTCH3L1519P receptor generates enhanced downstream signaling in a ligand-independent manner. Despite the enhanced signaling, the NOTCH3L1519P receptor is absent from the cell surface and instead accumulates in the endoplasmic reticulum. Furthermore, the localization of the NOTCH3L1519P receptor in the bipartite, heterodimeric state is altered, combined with avid secretion of the mutated extracellular domain from the cell. Chloroquine treatment strongly reduces the amount of secreted NOTCH3L1519P extracellular domain and decreases signaling. Finally, NOTCH3L1519P upregulates PDGFRB expression in fibroblasts, supporting a functional link between Notch and PDGF dysregulation in IMF. Collectively, our data define a NOTCH3–PDGFRB axis in IMF, in which an IMF-mutated NOTCH3 receptor elevates PDGFRB expression. The functional characterization of a ligand-independent gain-of-function NOTCH3 mutation is important for Notch therapy considerations for IMF, including strategies aimed at altering lysosome function.

scholarly journals Effect of Surface Topography Parameters on Friction and Wear of Random Rough Surface

Materials ◽  
2019 ◽  
Vol 12 (17) ◽  
pp. 2762 ◽  
Author(s):  
Ruimin Shi ◽  
Bukang Wang ◽  
Zhiwei Yan ◽  
Zongyan Wang ◽  
Lei Dong
Keyword(s):  
Surface Topography ◽  
Fluid Dynamic ◽  
Dynamic Pressure ◽  
Functional Characterization ◽  
Three Dimensional ◽  
Measuring System ◽  
Height Distribution ◽  
Worn Surface ◽  
The Relationship

In order to explore the relationship between the surface topography parameters and friction properties of a rough contact interface under fluid dynamic pressure lubrication conditions, friction experiments were carried out. The three-dimensional surface topography of specimens was measured and characterized with a profile microscopy measuring system and scanning electron microscope. The friction coefficient showed a trend of decreasing first and then increasing with the increase in some surface topography parameters at lower pressure, such as the surface height arithmetic mean Sa, surface height distribution kurtosis Sku, surface volume average volume Vvv, and surface center area average void volume Vvc, which are the ISO 25178 international standard parameters. The effects of surface topographic parameters on friction were analyzed and the wear mechanism of the worn surface was presented. The wear characteristics of the samples were mainly characterized as strain fatigue, grinding, and scraping. The results provide a theoretical basis for the functional characterization of surface topography.

In-vitro folding and functional characterization of the extracellular domain of G protein-coupled receptors

2002 ◽  
Vol 30 (3) ◽  
pp. A60-A60
Author(s):  
A. Kuntzsch ◽  
U. Grauschopf ◽  
A. Bazarsuren ◽  
K. Wenig ◽  
H. Lilie ◽  
...  
Keyword(s):  
G Protein ◽  
Functional Characterization ◽  
Extracellular Domain ◽  
G Protein Coupled Receptors ◽  
G Protein Coupled

scholarly journals Identification of diverse targets of arginine phosphorylation in Mycolicibacterium smegmatis by shotgun proteomics

2021 ◽  
Author(s):  
Emmanuel C Ogbonna ◽  
Karl R Schmitz
Keyword(s):  
Quality Control ◽  
Drug Targets ◽  
Functional Characterization ◽  
Shotgun Proteomics ◽  
Multidrug Resistant ◽  
Phosphopeptide Enrichment ◽  
Independent Manner ◽  
New Evidence ◽  
Lines Of Evidence

Tuberculosis is a leading cause of worldwide infectious mortality. The prevalence of multidrug-resistant Mycobacterium tuberculosis (Mtb) infections drives an urgent need to exploit new drug targets. One such target is the ATP-dependent protease ClpC1P1P2, which is strictly essential for viability. However, few proteolytic substrates of mycobacterial ClpC1P1P2 have been identified to date. Recent studies in Bacillus subtilis have shown that the orthologous ClpCP protease recognizes proteolytic substrates bearing post-translational arginine phosphorylation. Several lines of evidence suggest that ClpC1P1P2 similarly recognizes phosphoarginine-bearing proteins, but the existence of phosphoarginine modifications in mycobacteria has remained in question. Here, we confirm the presence of post-translational phosphoarginine modifications in Mycolicibacterium smegmatis (Msm), a nonpathogenic surrogate of Mtb. Using a phosphopeptide enrichment workflow coupled with shotgun phosphoproteomics, we identify arginine phosphosites on a diverse collection of targets within the Msm proteome. Physicochemical and functional characterization of targets suggest that arginine phosphorylation is applied in a sequence-independent manner as part of a proteome-wide quality control pathway. Our findings provide new evidence supporting the existence of phosphoarginine-mediated proteolysis by ClpC1P1P2 in mycobacteria and other actinobacterial species.

Prokaryotic Expression and Functional Characterization of the 19 kDa Protein in Balanus albicostatus Cement

2013 ◽  
Vol 461 ◽  
pp. 445-450
Author(s):  
Chao Liang ◽  
Yun Qiu Li ◽  
Bi Ru Hu ◽  
Wen Jian Wu
Keyword(s):  
Amino Acid ◽  
Molecular Design ◽  
Functional Characterization ◽  
Blue Staining ◽  
Amino Acid Residues ◽  
Adhesive Ability ◽  
Solid Liquid ◽  
Dental Applications ◽  
The Relationship

Barnacle is a unique sessile crustacean, which produces a multi-protein complex historically called barnacle cement to attach to diverse immersed materials permanently. The proteinaceous cement exhibits powerful adhesive property and special waterproof capability to cure at solid-liquid boundaries, which makes it ideal biomaterial for technical, medical and dental applications. It has been proved that a 19 kDa protein component, termed cp-19k in the cement plays a key role in surface coupling during underwater attachment. To verify whether the bacterial recombinant 19 kDa protein retains the adhesive ability, we cloned and sequenced the Bacp-19k gene in Balanus albicostatus. It encodes 173 amino acid residues, with seven biased ones, Thr, Lys, Gly, Ala, Val, Ser and Leu, comprising about 80% of the total. Two amino acid substitutions (F69L, I106L) were discovered in Bacp-19k due to the polymorphisms in barnacle cp-19ks, compared with the submitted one (GenBank: AB242295.1). Recombinant Bacp-19k was highly expressed in host strain Escherichia coli BL21 (DE3) and purified by affinity chromatography. Adsorption of recombinant Bacp-19k to glass substrata was examined by Coomassie brilliant blue staining. Future study will reveal the relationship between specific structures and functions for molecular design of novel biomimetic underwater adhesives.

scholarly journals Functional Characterization of Olfactory Receptors in the Thyroid Gland

2021 ◽  
Vol 12 ◽  
Author(s):  
Daniel Weidinger ◽  
Nikolina Jovancevic ◽  
Denise Zwanziger ◽  
Sarah Theurer ◽  
Judith Hönes ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Olfactory Receptors ◽  
Malignant Cell ◽  
Human Thyroid ◽  
Downstream Signaling ◽  
Cascade Analysis ◽  
Proliferation And Invasion ◽  
Specific Ligand ◽  
Stimulation Of

Olfactory receptors (ORs) are almost ubiquitously expressed in the human body. However, information about their functions in these tissues is lacking. To date, no functional characterization of expressed ORs in the human thyroid has been performed. In this study, we detected and compared the expression of OR2H2 and OR2W3 in healthy and malignant cell lines and their corresponding tissues, respectively. We demonstrated that stimulation of ORs by their specific ligand resulted in a transient increase in intracellular calcium and cAMP concentrations. In the case of OR2H2, the downstream signaling cascade analysis revealed that adenylate cyclase (AC) and phosphoinositide phospholipase C (PLC) were involved. Furthermore, OR2H2 and OR2W3 activation affected migration, proliferation, and invasion. These are the first insights that ORs influence physiology-relevant processes in the healthy and malignant thyroid.

scholarly journals Analysis of Secreted Proteins and Potential Virulence via the ICEs-Mediated Pathway of the Foodborne Pathogen Vibrio parahaemolyticus

2021 ◽  
Vol 12 ◽  
Author(s):  
Yu He ◽  
Shuai Wang ◽  
Kaiwen Wang ◽  
Jinwei Zhou ◽  
Zhi Han ◽  
...  
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Biochemical Characterization ◽  
Functional Characterization ◽  
Foodborne Pathogen ◽  
Secreted Proteins ◽  
Secretion Systems ◽  
Two Dimensional Gel Electrophoresis ◽  
Integrative And Conjugative Elements ◽  
The Relationship

Vibrio parahaemolyticus uses bacterial secretion systems and integrative and conjugative elements (ICEs) to induce various diseases and to adapt to harsh environments, respectively. Information pertaining to the identity of secreted proteins and functional characterization of ICEs has been previously reported, but the relationship between these elements remains unclear. Herein we investigated secreted proteins of V. parahaemolyticus strains JHY20 and JHY20△ICE using two-dimensional gel electrophoresis and LC-MS/MS, which led to the identification of an ICE-associated secreted protein – dihydrolipoamide dehydrogenase (DLDH). Considering the data related to its physical and biochemical characterization, we predicted that DLDH is a novel immunogenic protein and associated with virulence in JHY20. Our findings indicate a potential relationship between ICE-associated transport and secreted proteins and shed light on the function of such transport mechanisms. We believe that our data should enhance our understanding of mobile genetic elements.

scholarly journals Cryo-EM structures and functional characterization of the lipid scramblase TMEM16F

2018 ◽  
Author(s):  
Carolina Alvadia ◽  
Novandy K. Lim ◽  
Vanessa Clerico Mosina ◽  
Gert T. Oostergetel ◽  
Raimund Dutzler ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Functional Data ◽  
Functional Characterization ◽  
Anion Channel ◽  
Bound State ◽  
Striking Similarity ◽  
Common Mechanism ◽  
Protein Conformations ◽  
The Relationship

SUMMARYThe lipid scramblase TMEM16F initiates blood coagulation by catalyzing the exposure of phosphatidylserine in platelets. The protein is part of a family of membrane proteins, which encompasses calcium-activated channels for ions and lipids. Here, we reveal features of TMEM16F that underlie its function as lipid scramblase and ion channel. The cryo-EM structures of TMEM16F in Ca2+-bound and Ca2+-free states display a striking similarity to the scrambling-incompetent anion channel TMEM16A, yet with distinct differences in the catalytic site and in the conformational changes upon activation. In conjunction with functional data, we demonstrate the relationship between ion conduction and lipid scrambling. Although activated by a common mechanism, which likely resembles an equivalent process defined in the homologue nhTMEM16, both functions appear to be mediated by alternate protein conformations, which are at equilibrium in the ligand-bound state.

Molecular cloning and functional characterization of the porcine extracellular domain of Receptor Activator of NF-κB Ligand (sRANKL)

Gene ◽  
2012 ◽  
Vol 492 (1) ◽  
pp. 296-304 ◽  
Author(s):  
Wolfgang Böcker ◽  
Tamara Radic ◽  
Veronika Schönitzer ◽  
Florian Haasters ◽  
Wolf Mutschler ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Functional Characterization ◽  
Extracellular Domain ◽  
Receptor Activator

scholarly journals Functional Characterization of a Lassa Virus Fusion Inhibitors Adaptive Mutant

2020 ◽  
Author(s):  
Jiao Guo ◽  
Guangshun Zhang ◽  
Yang Liu ◽  
Junyuan Cao ◽  
Mengmeng Zhang ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Vaccine Design ◽  
Cross Resistance ◽  
Lassa Virus ◽  
Viral Growth ◽  
Glycoprotein Complex ◽  
Fusion Inhibitors ◽  
Tm Domain ◽  
The Relationship

ABSTRACTLassa virus (LASV) glycoprotein complex (GPC) contains retained stable-signal peptide (SSP), GP1, and GP2. SSP interacts with GP2 and provides an interface targeted by numerous fusion inhibitors. Serially passaging of LASV with inhibitors allowed some adaptive mutants to be obtained of which most had mutations located in the transmembrane (TM) domain of GP2. In the current study, we focused on the F446L mutant, which is reported to confer resistance to ST-series inhibitors. We found that F446L conferred cross-resistance to structurally distinct inhibitors. Furthermore, F446L increased the fusion activities of LASV and Mopeia virus GPC, elevating the pH threshold for fusion of LASV and promoting fusion of MOPV at neutral pH. F446L exerted little effect on the pseudotype viral growth profile or thermostability. By introducing other residues to the conserved F446 locus, it was found that this site was less compatible with a similar tyrosine residue and was intolerable to charged residues. These results help characterize the fusion inhibitor target located in the TM domain of GP2, which should be useful for drug and vaccine design.IMPORTANCEThe LASV SSP-GP2 interface provides an Achilles heel that is targeted by numerous inhibitors. However, the emergence of resistant viruses is a major concern for direct antiviral drugs. In this study, we investigated the F446L mutant located in the GPC TM domain to determine the relationship between drug resistance, membrane fusion activity, viral growth kinetics, and thermostability. These results will be helpful in monitoring drug-resistant variants, as well as the advancement of drug and vaccine design.

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