Online multi-channel microfluidic chip-mass spectrometry and its application for quantifying noncovalent protein–protein interactions

The Analyst ◽  
2015 ◽  
Vol 140 (5) ◽  
pp. 1551-1554 ◽  
Author(s):  
Wu Liu ◽  
Qiushui Chen ◽  
Xuexia Lin ◽  
Jin-Ming Lin
Keyword(s):  
Mass Spectrometry ◽  
Protein Interactions ◽  
Concanavalin A ◽  
Microfluidic Chip ◽  
Protein Protein Interactions ◽  
Paper Spray ◽  
Paper Spray Ionization ◽  
Noncovalent Protein

An online chip-MS platform interfaced by paper spray ionization has been developed in an automatic manner for multi-channel microfluidics, as demonstrated by the quantitation of dimer–tetramer interaction of concanavalin A.

scholarly journals PIPINO: A Software Package to Facilitate the Identification of Protein-Protein Interactions from Affinity Purification Mass Spectrometry Data

2016 ◽  
Vol 2016 ◽  
pp. 1-13
Author(s):  
Stefan Kalkhof ◽  
Stefan Schildbach ◽  
Conny Blumert ◽  
Friedemann Horn ◽  
Martin von Bergen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Interactions ◽  
Isotope Labeling ◽  
Affinity Purification ◽  
Interaction Network ◽  
Mass Spectrometry Data ◽  
Protein Protein Interactions ◽  
Protein Protein Interaction ◽  
Binding Behavior ◽  
Bona Fide

The functionality of most proteins is regulated by protein-protein interactions. Hence, the comprehensive characterization of the interactome is the next milestone on the path to understand the biochemistry of the cell. A powerful method to detect protein-protein interactions is a combination of coimmunoprecipitation or affinity purification with quantitative mass spectrometry. Nevertheless, both methods tend to precipitate a high number of background proteins due to nonspecific interactions. To address this challenge the software Protein-Protein-Interaction-Optimizer (PIPINO) was developed to perform an automated data analysis, to facilitate the selection of bona fide binding partners, and to compare the dynamic of interaction networks. In this study we investigated the STAT1 interaction network and its activation dependent dynamics. Stable isotope labeling by amino acids in cell culture (SILAC) was applied to analyze the STAT1 interactome after streptavidin pull-down of biotagged STAT1 from human embryonic kidney 293T cells with and without activation. Starting from more than 2,000 captured proteins 30 potential STAT1 interaction partners were extracted. Interestingly, more than 50% of these were already reported or predicted to bind STAT1. Furthermore, 16 proteins were found to affect the binding behavior depending on STAT1 phosphorylation such as STAT3 or the importin subunits alpha 1 and alpha 6.

Paper Spray Ionization of Noncovalent Protein Complexes

2014 ◽  
Vol 86 (3) ◽  
pp. 1342-1346 ◽  
Author(s):  
Yun Zhang ◽  
Yue Ju ◽  
Chengsi Huang ◽  
Vicki H. Wysocki
Keyword(s):  
Protein Complexes ◽  
Paper Spray ◽  
Paper Spray Ionization ◽  
Noncovalent Protein

scholarly journals Mass spectrometry-based cross-linking study shows that the Psb28 protein binds to cytochrome b559 in Photosystem II

2017 ◽  
Vol 114 (9) ◽  
pp. 2224-2229 ◽  
Author(s):  
Daniel A. Weisz ◽  
Haijun Liu ◽  
Hao Zhang ◽  
Sundarapandian Thangapandian ◽  
Emad Tajkhorshid ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Photosystem Ii ◽  
Protein Interactions ◽  
Protein Complexes ◽  
Protein Docking ◽  
Cross Linking ◽  
Protein Protein Interactions ◽  
Cytochrome B559 ◽  
Reaction Center Complex ◽  
Assembly Intermediate

Photosystem II (PSII), a large pigment protein complex, undergoes rapid turnover under natural conditions. During assembly of PSII, oxidative damage to vulnerable assembly intermediate complexes must be prevented. Psb28, the only cytoplasmic extrinsic protein in PSII, protects the RC47 assembly intermediate of PSII and assists its efficient conversion into functional PSII. Its role is particularly important under stress conditions when PSII damage occurs frequently. Psb28 is not found, however, in any PSII crystal structure, and its structural location has remained unknown. In this study, we used chemical cross-linking combined with mass spectrometry to capture the transient interaction of Psb28 with PSII. We detected three cross-links between Psb28 and the α- and β-subunits of cytochrome b559, an essential component of the PSII reaction-center complex. These distance restraints enable us to position Psb28 on the cytosolic surface of PSII directly above cytochrome b559, in close proximity to the QB site. Protein–protein docking results also support Psb28 binding in this region. Determination of the Psb28 binding site and other biochemical evidence allow us to propose a mechanism by which Psb28 exerts its protective effect on the RC47 intermediate. This study also shows that isotope-encoded cross-linking with the “mass tags” selection criteria allows confident identification of more cross-linked peptides in PSII than has been previously reported. This approach thus holds promise to identify other transient protein–protein interactions in membrane protein complexes.

Direct analysis of herbicides by paper spray ionization mass spectrometry

2015 ◽  
Vol 7 (23) ◽  
pp. 9808-9816 ◽  
Author(s):  
Steven L. Reeber ◽  
Sneha Gadi ◽  
Sung-Ben Huang ◽  
Gary L. Glish
Keyword(s):  
Mass Spectrometry ◽  
Environmental Samples ◽  
Direct Analysis ◽  
Mass Spectrometric ◽  
Mass Spectrometric Analysis ◽  
Spectrometric Analysis ◽  
Ionization Mass ◽  
Paper Spray ◽  
Paper Spray Ionization ◽  
Rapid Mass

Paper spray ionization enables the rapid mass spectrometric analysis of environmental samples without the use of chromatography or sample cleanup techniques.

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