A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

10.3791/58582 ◽  
2018 ◽  
Author(s):  
Valentin Dunsing ◽  
Salvatore Chiantia
Keyword(s):  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Fluorescence Fluctuation Spectroscopy ◽  
Cell Contacts ◽  
Cell Cell

Characterization of the interactions of alpha-catenin with alpha-actinin and beta-catenin/plakoglobin

1997 ◽  
Vol 110 (8) ◽  
pp. 1013-1022 ◽  
Author(s):  
J.E. Nieset ◽  
A.R. Redfield ◽  
F. Jin ◽  
K.A. Knudsen ◽  
K.R. Johnson ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Protein Interactions ◽  
Beta Catenin ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Calcium Dependent ◽  
Cytoplasmic Proteins ◽  
Dependent Cell ◽  
Two Hybrid ◽  
Cell Cell

Cadherins are calcium-dependent, cell surface glycoproteins involved in cell-cell adhesion. To function in cell-cell adhesion, the transmembrane cadherin molecule must be associated with the cytoskeleton via cytoplasmic proteins known as catenins. Three catenins, alpha-catenin, beta-catenin and gamma-catenin (also known as plakoglobin), have been identified. beta-catenin or plakoglobin is associated directly with the cadherin; alpha-catenin binds to beta-catenin/plakoglobin and serves to link the cadherin/catenin complex to the actin cytoskeleton. The domains on the cadherin and betacatenin/plakoglobin that are responsible for protein-protein interactions have been mapped. However, little is known about the molecular interactions between alpha-catenin and beta-catenin/plakoglobin or about the interactions between alpha-catenin and the cytoskeleton. In this study we have used the yeast two-hybrid system to map the domains on alpha-catenin that allow it to associate with beta-catenin/plakoglobin and with alpha-actinin. We also identify a region on alpha-actinin that is responsible for its interaction with alpha-catenin. The yeast two-hybrid data were confirmed with biochemical studies.

scholarly journals Residues in a Highly Conserved Claudin-1 Motif Are Required for Hepatitis C Virus Entry and Mediate the Formation of Cell-Cell Contacts

2009 ◽  
Vol 83 (11) ◽  
pp. 5477-5484 ◽  
Author(s):  
Lisa Cukierman ◽  
Laurent Meertens ◽  
Claire Bertaux ◽  
Francis Kajumo ◽  
Tatjana Dragic
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Protein Interactions ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Membrane Domains ◽  
Cell Contacts ◽  
Functional Roles ◽  
Critical Residues ◽  
Cell Cell

ABSTRACT Claudin-1, a component of tight junctions between liver hepatocytes, is a hepatitis C virus (HCV) late-stage entry cofactor. To investigate the structural and functional roles of various claudin-1 domains in HCV entry, we applied a mutagenesis strategy. Putative functional intracellular claudin-1 domains were not important. However, we identified seven novel residues in the first extracellular loop that are critical for entry of HCV isolates drawn from six different subtypes. Most of the critical residues belong to the highly conserved claudin motif W30-GLW51-C54-C64. Alanine substitutions of these residues did not impair claudin-1 cell surface expression or lateral protein interactions within the plasma membrane, including claudin-1-claudin-1 and claudin-1-CD81 interactions. However, these mutants no longer localized to cell-cell contacts. Based on our observations, we propose that cell-cell contacts formed by claudin-1 may generate specialized membrane domains that are amenable to HCV entry.

scholarly journals Cellinker: a platform of ligand–receptor interactions for intercellular communication analysis

2021 ◽  
Author(s):  
Yang Zhang ◽  
Tianyuan Liu ◽  
Jing Wang ◽  
Bohao Zou ◽  
Le Li ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Intercellular Communication ◽  
Rapid Development ◽  
Cell Communication ◽  
Supplementary Information ◽  
Protein Protein Interactions ◽  
Communication Analysis ◽  
Mediate Cell ◽  
Intercellular Communications ◽  
Cell Cell

Abstract Motivation Ligand–receptor (L–R) interactions mediate cell adhesion, recognition and communication and play essential roles in physiological and pathological signaling. With the rapid development of single-cell RNA sequencing (scRNA-seq) technologies, systematically decoding the intercellular communication network involving L–R interactions has become a focus of research. Therefore, construction of a comprehensive, high-confidence and well-organized resource to retrieve L–R interactions in order to study the functional effects of cell–cell communications would be of great value. Results In this study, we developed Cellinker, a platform of literature-supported L–R interactions that play roles in cell–cell communication. We aimed to provide a useful platform for studies on cell–cell communication mediated by L–R interactions. The current version of Cellinker documents over 3700 human and 3200 mouse L–R protein–protein interactions (PPIs) and embeds a practical and convenient webserver with which researchers can decode intercellular communications based on scRNA-seq data. And over 400 endogenous small molecule (sMOL) related L–R interactions were collected as well. Moreover, to help with research on coronavirus (CoV) infection, Cellinker collects information on 16L–R PPIs involved in CoV–human interactions (including 12L–R PPIs involved in SARS-CoV-2 infection). In summary, Cellinker provides a user-friendly interface for querying, browsing and visualizing L–R interactions as well as a practical and convenient web tool for inferring intercellular communications based on scRNA-seq data. We believe this platform could promote intercellular communication research and accelerate the development of related algorithms for scRNA-seq studies. Availability and implementation Cellinker is available at http://www.rna-society.org/cellinker/ Supplementary information Supplementary data are available at Bioinformatics online.

CellTalkDB: a manually curated database of ligand–receptor interactions in humans and mice

2020 ◽  
Author(s):  
Xin Shao ◽  
Jie Liao ◽  
Chengyu Li ◽  
Xiaoyan Lu ◽  
Junyun Cheng ◽  
...  
Keyword(s):  
Single Cell ◽  
Protein Interactions ◽  
Cell Communication ◽  
Systematic Investigation ◽  
Protein Protein Interactions ◽  
Biological Phenomena ◽  
Type Composition ◽  
Dependent Cell ◽  
Multicellular Organisms ◽  
Cell Cell

Abstract Cell–cell communications in multicellular organisms generally involve secreted ligand–receptor (LR) interactions, which is vital for various biological phenomena. Recent advancements in single-cell RNA sequencing (scRNA-seq) have effectively resolved cellular phenotypic heterogeneity and the cell-type composition of complex tissues, facilitating the systematic investigation of cell–cell communications at single-cell resolution. However, assessment of chemical-signal-dependent cell–cell communication through scRNA-seq relies heavily on prior knowledge of LR interaction pairs. We constructed CellTalkDB (http://tcm.zju.edu.cn/celltalkdb), a manually curated comprehensive database of LR interaction pairs in humans and mice comprising 3398 human LR pairs and 2033 mouse LR pairs, through text mining and manual verification of known protein–protein interactions using the STRING database, with literature-supported evidence for each pair. Compared with SingleCellSignalR, the largest LR-pair resource, CellTalkDB includes not only 2033 mouse LR pairs but also 377 additional human LR pairs. In conclusion, the data on human and mouse LR pairs contained in CellTalkDB could help to further the inference and understanding of the LR-interaction-based cell–cell communications, which might provide new insights into the mechanism underlying biological processes.

scholarly journals Heparan Sulfate Biosynthesis and Sulfation Profiles as Modulators of Cancer Signalling and Progression

2021 ◽  
Vol 11 ◽  
Author(s):  
Catarina Marques ◽  
Celso A. Reis ◽  
Romain R. Vivès ◽  
Ana Magalhães
Keyword(s):  
Heparan Sulfate ◽  
Protein Interactions ◽  
Cell Signalling ◽  
Structural Diversity ◽  
Structural Features ◽  
Protein Protein Interactions ◽  
Cellular Events ◽  
And Function ◽  
The Impact ◽  
Cell Cell

Heparan Sulfate Proteoglycans (HSPGs) are important cell surface and Extracellular Matrix (ECM) maestros involved in the orchestration of multiple cellular events in physiology and pathology. These glycoconjugates bind to various bioactive proteins via their Heparan Sulfate (HS) chains, but also through the protein backbone, and function as scaffolds for protein-protein interactions, modulating extracellular ligand gradients, cell signalling networks and cell-cell/cell-ECM interactions. The structural features of HS chains, including length and sulfation patterns, are crucial for the biological roles displayed by HSPGs, as these features determine HS chains binding affinities and selectivity. The large HS structural diversity results from a tightly controlled biosynthetic pathway that is differently regulated in different organs, stages of development and pathologies, including cancer. This review addresses the regulatory mechanisms underlying HS biosynthesis, with a particular focus on the catalytic activity of the enzymes responsible for HS glycan sequences and sulfation motifs, namely D-Glucuronyl C5-Epimerase, N- and O-Sulfotransferases. Moreover, we provide insights on the impact of different HS structural epitopes over HSPG-protein interactions and cell signalling, as well as on the effects of deregulated expression of HS modifying enzymes in the development and progression of cancer. Finally, we discuss the clinical potential of HS biosynthetic enzymes as novel targets for therapy, and highlight the importance of developing new HS-based tools for better patients’ stratification and cancer treatment.

Protein-protein interactions of the p53 family with the Bcl-2 family in hepatocellular carcinoma

2011 ◽  
Vol 49 (08) ◽  
Author(s):  
LC König ◽  
M Meinhard ◽  
C Sandig ◽  
MH Bender ◽  
A Lovas ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Protein Interactions ◽  
Protein Protein Interactions ◽  
P53 Family

Partial Purification of a Specific Plasminogen Activator from Porcine Parotid Glands

1974 ◽  
Vol 31 (03) ◽  
pp. 403-414 ◽  
Author(s):  
Terence Cartwright
Keyword(s):  
Nucleic Acid ◽  
Salivary Glands ◽  
Plasminogen Activator ◽  
Protein Interactions ◽  
Partial Purification ◽  
Protein Protein Interactions ◽  
Parotid Glands ◽  
Two Stage ◽  
Preliminary Characterization ◽  
Specific Inhibitors

SummaryA method is described for the extraction with buffers of near physiological pH of a plasminogen activator from porcine salivary glands. Substantial purification of the activator was achieved although this was to some extent complicated by concomitant extraction of nucleic acid from the glands. Preliminary characterization experiments using specific inhibitors suggested that the activator functioned by a similar mechanism to that proposed for urokinase, but with some important kinetic differences in two-stage assay systems. The lack of reactivity of the pig gland enzyme in these systems might be related to the tendency to protein-protein interactions observed with this material.

Characterization of N-cadherin-containing cell-cell contacts of the liver

2006 ◽  
Vol 44 (01) ◽  
Author(s):  
BK Straub ◽  
J Boda-Heggemann ◽  
UF Pape ◽  
C Grund ◽  
E Specht-Delius ◽  
...  
Keyword(s):  
Cell Contacts ◽  
Cell Cell
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