scholarly journals Label-free quantitative proteomics trends for protein–protein interactions

2013 ◽  
Vol 81 ◽  
pp. 91-101 ◽  
Author(s):  
Stephen Tate ◽  
Brett Larsen ◽  
Ron Bonner ◽  
Anne-Claude Gingras
Keyword(s):  
Protein Interactions ◽  
Quantitative Proteomics ◽  
Label Free ◽  
Protein Protein Interactions

scholarly journals Analysis of Zika virus capsid-Aedes aegypti mosquito interactome reveals pro-viral host factors critical for establishing infection

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Rommel J. Gestuveo ◽  
Jamie Royle ◽  
Claire L. Donald ◽  
Douglas J. Lamont ◽  
Edward C. Hutchinson ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Protein Interactions ◽  
Zika Virus ◽  
Label Free ◽  
Protein Protein Interactions ◽  
Zikv Infection ◽  
Ubiquitin Protein Ligase ◽  
Mosquito Cells ◽  
Endoplasmic Reticulum Protein ◽  
Arboviral Diseases

AbstractThe escalating global prevalence of arboviral diseases emphasizes the need to improve our understanding of their biology. Research in this area has been hindered by the lack of molecular tools for studying virus-mosquito interactions. Here, we develop an Aedes aegypti cell line which stably expresses Zika virus (ZIKV) capsid proteins in order to study virus-vector protein-protein interactions through quantitative label-free proteomics. We identify 157 interactors and show that eight have potentially pro-viral activity during ZIKV infection in mosquito cells. Notably, silencing of transitional endoplasmic reticulum protein TER94 prevents ZIKV capsid degradation and significantly reduces viral replication. Similar results are observed if the TER94 ortholog (VCP) functioning is blocked with inhibitors in human cells. In addition, we show that an E3 ubiquitin-protein ligase, UBR5, mediates the interaction between TER94 and ZIKV capsid. Our study demonstrates a pro-viral function for TER94/VCP during ZIKV infection that is conserved between human and mosquito cells.

scholarly journals Label-free methods for optical in vitro characterization of protein–protein interactions

2021 ◽  
Author(s):  
Fabian Soltermann ◽  
Weston B. Struwe ◽  
Philipp Kukura
Keyword(s):  
Protein Interactions ◽  
Label Free ◽  
Protein Protein Interactions ◽  
Cellular Processes ◽  
P Protein ◽  
In Vitro Characterization ◽  
And Function

Protein–protein interactions are involved in the regulation and function of the majority of cellular processes.

Effect of Intestinal Flora Clearance on Liver Proteomics in Mice

2019 ◽  
Vol 16 (3) ◽  
pp. 199-209
Author(s):  
Zhenghu Jia ◽  
Hui Liu ◽  
Mei Song ◽  
Chengmao Yang ◽  
Yapu Zhao ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Concanavalin A ◽  
Intestinal Flora ◽  
Label Free ◽  
Protein Protein Interactions ◽  
Rt Pcr ◽  
Proteome Level ◽  
And Control ◽  
The Relationship ◽  
Free Quantification

Background: Intestinal flora dynamically affects the host&#039;s systemic immune system. Liver is one of the organs that may be affected by intestinal microbiota. </P><P> Materials and Methods: In this study, we aimed to identify proteome level differences between liver tissue from mice cleared intestinal flora and control using tandem mass spectrometry (LC-MS/MS) and label free quantification. Additionally, protein-protein interactions were mapped by STRING, and also, the enrichment of inflammation-related signaling pathways and biological processes was identified using GO and IPA network system. RT-PCR and Western blot were used for validation of the proteomics findings. Results: Our study demonstrated that mice with cleared intestinal flora exhibited decreased sensitivity to Concanavalin A induced acute hepatitis. 324 Proteins in liver were differently expressed after intestinal flora clearance for one week while 210 proteins were differently expressed after intestinal flora clearance for two weeks. Furthermore, five of the identified proteins were validated by western blotting and further investigated by semi-quantitative RT-PCR. Conclusion: Our results showed that intestinal flora clearance in mice could reduce sensitivity to Concanavalin A induced liver injury and influence the expression of proteins in liver, which provides a clue for studying the relationship between gut bacteria and Concanavalin A induced hepatitis.

Label-Free Real-Time Microarray Imaging of Cancer Protein–Protein Interactions and Their Inhibition by Small Molecules

2016 ◽  
Vol 88 (6) ◽  
pp. 3130-3135 ◽  
Author(s):  
Charuksha Walgama ◽  
Zainab H. Al Mubarak ◽  
Bing Zhang ◽  
Mayowa Akinwale ◽  
Anuruddha Pathiranage ◽  
...  
Keyword(s):  
Small Molecules ◽  
Real Time ◽  
Protein Interactions ◽  
Label Free ◽  
Protein Protein Interactions

Label-free quantitative proteomic analysis of the YAP/TAZ interactome

2014 ◽  
Vol 306 (9) ◽  
pp. C805-C818 ◽  
Author(s):  
Priyanka Kohli ◽  
Malte P. Bartram ◽  
Sandra Habbig ◽  
Caroline Pahmeyer ◽  
Tobias Lamkemeyer ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Interactions ◽  
Affinity Purification ◽  
Single Copy ◽  
Single Step ◽  
Mass Spectrometry Analysis ◽  
Single Shot ◽  
Label Free ◽  
Protein Protein Interactions ◽  
Spectrometry Analysis

The function of an individual protein is typically defined by protein-protein interactions orchestrating the formation of large complexes critical for a wide variety of biological processes. Over the last decade the analysis of purified protein complexes by mass spectrometry became a key technique to identify protein-protein interactions. We present a fast and straightforward approach for analyses of interacting proteins combining a Flp-in single-copy cellular integration system and single-step affinity purification with single-shot mass spectrometry analysis. We applied this protocol to the analysis of the YAP and TAZ interactome. YAP and TAZ are the downstream effectors of the mammalian Hippo tumor suppressor pathway. Our study provides comprehensive interactomes for both YAP and TAZ and does not only confirm the majority of previously described interactors but, strikingly, revealed uncharacterized interaction partners that affect YAP/TAZ TEAD-dependent transcription. Among these newly identified candidates are Rassf8, thymopoetin, and the transcription factors CCAAT/enhancer-binding protein (C/EBP)β/δ and core-binding factor subunit β (Cbfb). In addition, our data allowed insights into complex stoichiometry and uncovered discrepancies between the YAP and TAZ interactomes. Taken together, the stringent approach presented here could help to significantly sharpen the understanding of protein-protein networks.

Sign in / Sign up

Export Citation Format

Share Document