scholarly journals Subcellular Localization of Connexin 26 in Cardiomyocytes and in Cardiomyocyte-Derived Extracellular Vesicles

Molecules ◽  
2021 ◽  
Vol 26 (21) ◽  
pp. 6726
Author(s):  
Alessandra Falleni ◽  
Stefania Moscato ◽  
Antonietta R. M. Sabbatini ◽  
Margherita Bernardeschi ◽  
Francesco Bianchi ◽  
...  
Keyword(s):  
Subcellular Localization ◽  
Extracellular Vesicles ◽  
Heart Function ◽  
Functional Characterization ◽  
Tissue Homeostasis ◽  
Connexin 26 ◽  
Cardiac Cell ◽  
Intercalated Discs ◽  
Membrane Spanning ◽  
Subcellular Level

Connexins (Cxs) are a family of membrane-spanning proteins, expressed in vertebrates and named according to their molecular weight. They are involved in tissue homeostasis, and they function by acting at several communication levels. Cardiac Cxs are responsible for regular heart function and, among them, Cx26 and Cx43 are widely expressed throughout the heart. Cx26 is present in vessels, as well as in cardiomyocytes, and its localization is scattered all over the cell aside from at the intercalated discs as is the case for the other cardiac Cxs. However, having been found in cardiomyocytes only recently, both its subcellular localization and its functional characterization in cardiomyocytes remain poorly understood. Therefore, in this study we aimed to obtain further data on the localization of Cx26 at the subcellular level. Our TEM immunogold analyses were performed on rat heart ventricles and differentiated H9c2 cardiac cell sections as well as on differentiated H9c2 derived extracellular vesicles. The results confirmed the absence of Cx26 at intercalated discs and showed the presence of Cx26 at the level of different subcellular compartments. The peculiar localization at the level of extracellular vesicles suggested a specific role for cardiac Cx26 in inter-cellular communication in an independent gap junction manner.

scholarly journals Proteomic, transcriptomic and functional characterization of human astrocyte‐derived extracellular vesicles upon inflammatory activation

2020 ◽  
Vol 16 (S2) ◽  
Author(s):  
Yang You ◽  
Kathleen Borgmann ◽  
VenkataViswa Edara ◽  
Satomi Stacy ◽  
Anuja Ghorpade ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Functional Characterization ◽  
Human Astrocyte ◽  
Inflammatory Activation

scholarly journals Localization and function of a Plasmodium falciparum protein (PF3D7_1459400) during erythrocyte invasion

2020 ◽  
pp. 153537022096176
Author(s):  
Emmanuel Amlabu ◽  
Prince B Nyarko ◽  
Grace Opoku ◽  
Damata Ibrahim-Dey ◽  
Philip Ilani ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Subcellular Localization ◽  
Protein Expression ◽  
Functional Characterization ◽  
Blood Stage ◽  
Protein Subcellular Localization ◽  
Erythrocyte Invasion ◽  
Invasion Inhibition ◽  
Blood Stage Malaria ◽  
Peptide Antibodies

Nearly 60% of Plasmodium falciparum proteins are still uncharacterized and their functions are unknown. In this report, we carried out the functional characterization of a 45 kDa protein (PF3D7_1459400) and showed its potential as a target for blood stage malaria vaccine development. Analysis of protein subcellular localization, native protein expression profile, and erythrocyte invasion inhibition of both clinical and laboratory parasite strains by peptide antibodies suggest a functional role of PF3D7_1459400 protein during erythrocyte invasion. Also, immunoreactivity screens using synthetic peptides of the protein showed that adults resident in malaria endemic regions in Ghana have naturally acquired plasma antibodies against PF3D7_1459400 protein. Altogether, this study presents PF3D7_1459400 protein as a potential target for the development of peptide-based vaccine for blood-stage malaria. Impact statement Plasmodium falciparum malaria is a global health problem. Erythrocyte invasion by P. falciparum merozoites appears to be a promising target to curb malaria. We have identified and characterized a novel protein that is involved in erythrocyte invasion. Our data on protein subcellular localization, stage-specific protein expression pattern, and merozoite invasion inhibition by α-peptide antibodies suggest a role for PF3D7_1459400 protein during P. falciparum erythrocyte invasion. Even more, the human immunoepidemiology data present PF3D7_1459400 protein as an immunogenic antigen which could be further exploited for the development of new anti-infective therapy against malaria.

scholarly journals BARP suppresses voltage-gated calcium channel activity and Ca2+-evoked exocytosis

2014 ◽  
Vol 205 (2) ◽  
pp. 233-249 ◽  
Author(s):  
Pascal Béguin ◽  
Kazuaki Nagashima ◽  
Ramasubbu N. Mahalakshmi ◽  
Réjan Vigot ◽  
Atsuko Matsunaga ◽  
...  
Keyword(s):  
Subcellular Localization ◽  
Functional Characterization ◽  
Regulatory Protein ◽  
Binding Pocket ◽  
Neuroendocrine Cells ◽  
Cell Surface Expression ◽  
Channel Activity ◽  
Interaction Domain ◽  
Voltage Gated ◽  
Domain I

Voltage-gated calcium channels (VGCCs) are key regulators of cell signaling and Ca2+-dependent release of neurotransmitters and hormones. Understanding the mechanisms that inactivate VGCCs to prevent intracellular Ca2+ overload and govern their specific subcellular localization is of critical importance. We report the identification and functional characterization of VGCC β-anchoring and -regulatory protein (BARP), a previously uncharacterized integral membrane glycoprotein expressed in neuroendocrine cells and neurons. BARP interacts via two cytosolic domains (I and II) with all Cavβ subunit isoforms, affecting their subcellular localization and suppressing VGCC activity. Domain I interacts at the α1 interaction domain–binding pocket in Cavβ and interferes with the association between Cavβ and Cavα1. In the absence of domain I binding, BARP can form a ternary complex with Cavα1 and Cavβ via domain II. BARP does not affect cell surface expression of Cavα1 but inhibits Ca2+ channel activity at the plasma membrane, resulting in the inhibition of Ca2+-evoked exocytosis. Thus, BARP can modulate the localization of Cavβ and its association with the Cavα1 subunit to negatively regulate VGCC activity.

scholarly journals Identification of murine phosphodiesterase 5A isoforms and their functional characterization in HL-1 cardiac cell line

2017 ◽  
Vol 233 (1) ◽  
pp. 325-337 ◽  
Author(s):  
Federica Campolo ◽  
Alessandra Zevini ◽  
Silvia Cardarelli ◽  
Lucia Monaco ◽  
Federica Barbagallo ◽  
...  
Keyword(s):  
Cell Line ◽  
Functional Characterization ◽  
Cardiac Cell

Exosomes and extracellular vesicles: the path forward

2018 ◽  
Vol 62 (2) ◽  
pp. 119-124 ◽  
Author(s):  
Philip D. Stahl ◽  
Graça Raposo
Keyword(s):  
Extracellular Vesicles ◽  
Cell Biology ◽  
Biological Fluids ◽  
Therapeutic Agents ◽  
Tissue Homeostasis ◽  
The Past ◽  
And Function ◽  
Intercellular Communications ◽  
Biomedical Community

Over the course of the past several decades, the concept that extracellular vesicles, exosomes and microvesicles, operate as cellular “housekeepers” and as agents for communication between and among cells and tissues, has emerged into one of the most promising yet vexing problems facing the biomedical community. Already, extracellular vesicles from biological fluids are being used for diagnostic purposes and hopes abound for their use as therapeutic agents. However, the most basic mechanistic questions surrounding their biogenesis and function in cellular and tissue homeostasis remain largely unexplored. In this issue of Essays in Biochemistry, the rise of a new intercellular communications pathway is considered from many perspectives—cell biology, physiology, and pathophysiology.

scholarly journals Molecular Cloning, Functional Characterization, and Subcellular Localization of Soybean Nodule Dihydrolipoamide Reductase

2002 ◽  
Vol 128 (1) ◽  
pp. 300-313 ◽  
Author(s):  
Jose F. Moran ◽  
Zhaohui Sun ◽  
Gautam Sarath ◽  
Raúl Arredondo-Peter ◽  
Euan K. James ◽  
...  
Keyword(s):  
Subcellular Localization ◽  
Molecular Cloning ◽  
Functional Characterization ◽  
Soybean Nodule

Ion homeostasis, channels, and transporters: an update on cellular mechanisms

2004 ◽  
Vol 28 (4) ◽  
pp. 143-154 ◽  
Author(s):  
George R. Dubyak
Keyword(s):  
Ion Channels ◽  
Subcellular Localization ◽  
Ion Transport ◽  
Membrane Transport ◽  
Cell Biology ◽  
Functional Characterization ◽  
Transport Proteins ◽  
Transport Protein ◽  
Membrane Transport Proteins ◽  
Functional Analyses

The steady-state maintenance of highly asymmetric concentrations of the major inorganic cations and anions is a major function of both plasma membranes and the membranes of intracellular organelles. Homeostatic regulation of these ionic gradients is critical for most functions. Due to their charge, the movements of ions across biological membranes necessarily involves facilitation by intrinsic membrane transport proteins. The functional characterization and categorization of membrane transport proteins was a major focus of cell physiological research from the 1950s through the 1980s. On the basis of these functional analyses, ion transport proteins were broadly divided into two classes: channels and carrier-type transporters (which include exchangers, cotransporters, and ATP-driven ion pumps). Beginning in the mid-1980s, these functional analyses of ion transport and homeostasis were complemented by the cloning of genes encoding many ion channels and transporter proteins. Comparison of the predicted primary amino acid sequences and structures of functionally similar ion transport proteins facilitated their grouping within families and superfamilies of structurally related membrane proteins. Postgenomics research in ion transport biology increasingly involves two powerful approaches. One involves elucidation of the molecular structures, at the atomic level in some cases, of model ion transport proteins. The second uses the tools of cell biology to explore the cell-specific function or subcellular localization of ion transport proteins. This review will describe how these approaches have provided new, and sometimes surprising, insights regarding four major questions in current ion transporter research. 1) What are the fundamental differences between ion channels and ion transporters? 2) How does the interaction of an ion transport protein with so-called adapter proteins affect its subcellular localization or regulation by various intracellular signal transduction pathways? 3) How does the specific lipid composition of the local membrane microenvironment modulate the function of an ion transport protein? 4) How can the basic functional properties of a ubiquitously expressed ion transport protein vary depending on the cell type in which it is expressed?

scholarly journals Diverse populations of extracellular vesicles with opposite functions during herpes simplex virus 1 infection

2020 ◽  
Author(s):  
Christos Dogrammatzis ◽  
Shadia Saleh ◽  
Clayton Deighan ◽  
Maria Kalamvoki
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Extracellular Vesicles ◽  
Functional Characterization ◽  
Cell Communication ◽  
Antiviral Effect ◽  
Herpes Simplex Virus 1 ◽  
Infected Cells ◽  
Simplex Virus ◽  
Hsv 1

Extracellular vesicles (EVs) are released by all types of cells as a means of intercellular communication. Their significance lies in the fact that they can alter recipient cells functions, despite their limited capacity for cargo. We have previously demonstrated that herpes simplex virus 1 (HSV-1) infection influences the cargo and functions of EVs released by infected cells, and that these EVs negatively impact a subsequent HSV-1 infection. In the present study, we have implemented cutting-edge technologies to further characterize EVs released during HSV-1 infection. We identified distinct EV populations that were separable through a gradient approach. One population was positive for the tetraspanin CD63 and was distinct from EVs carrying components of the endosomal sorting complexes required for transport (ESCRT). Nanoparticle tracking analysis (NTA) combined with protein analysis indicated that the production of CD63+ EVs was selectively induced upon HSV-1 infection. The ExoView platform supported these data and suggested that the amount of CD63 per vesicle is higher upon infection. This platform also identified EV populations positive for other tetraspanins, including CD81 and CD9, whose abundance decreased upon HSV-1 infection. The STimulator of INterferon Genes (STING) was found in CD63+ EVs released during HSV-1 infection, while viral components were found in ESCRT+ EVs. Functional characterization of these EVs demonstrated that they have opposite effects on the infection, but the dominant effect was negative. Overall, we have identified the dominant population of EVs, and other EV populations produced during HSV-1 infection, and we have provided information about potential roles. Importance Extracellular vesicles mediate cell-to-cell communication and convey messages important for cell homeostasis. Pathways of EV biogenesis are often hijacked by pathogens to facilitate their dissemination and to establish a favorable microenvironment for the infection. We have previously shown that HSV-1 infection alters the cargo and functions of the released EVs, which negatively impact the infection. We have built upon our previous findings by developing procedures to separate EV populations from HSV-1 infected cells. We identified the major population of EVs released during infection, which carries the DNA sensor STING and has an antiviral effect. We also identified an EV population that carries selected viral proteins and has a pro-viral role. This is the first study to characterize EV populations during infection. These data indicate that the complex interactions between the virus and the host are extended to the extracellular environment and could impact HSV-1 dissemination and persistence in the host.

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