scholarly journals Sequential Peptide Affinity Purification System for the Systematic Isolation and Identification of Protein Complexes from Escherichia coli

2009 ◽  
pp. 373-400 ◽  
Author(s):  
Mohan Babu ◽  
Gareth Butl ◽  
Oxana Pogoutse ◽  
Joyce Li ◽  
Jack F. Greenblatt ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Complexes ◽  
Affinity Purification ◽  
Isolation And Identification ◽  
Purification System ◽  
Peptide Affinity

scholarly journals Optimization of Tandem Affinity Chromatography (TAP) for Effective Isolation and Identification of Protein Complexes Involved in Molecular Mechanism of Depression.

2021 ◽  
Author(s):  
Ewelina Fic ◽  
Agata Cieslik ◽  
Marta Dziedzicka-Wasylewska
Keyword(s):  
Molecular Mechanism ◽  
Depressive Disorders ◽  
Protein Complexes ◽  
Affinity Purification ◽  
Mitogen Activated Protein Kinase ◽  
World Health ◽  
Nucleic Acid Binding ◽  
Isolation And Identification ◽  
Mammalian Cell Lines ◽  
New Findings

Abstract According to the World Health Organization Report, depressive disorders affect about 10% of the population. The molecular mechanism of the pathogenesis of depression is still not well understood. The new findings point to phosphatases as potential targets for effective depression therapy. The aim of the project is the development of method that would enable the identification of Mitogen-Activated Protein Kinase Phosphatase-1 (MKP-1) protein partners using proteomic techniques. The research was carried out using the PC12 cell line, often used as a model for neurobiological research. The use of the procedure for efficient purification of protein complexes – Tandem Affinity Purification (TAP) will facilitate the identification of proteins interacting with MKP-1, a potential goal of effective antidepressant therapy. The presented protocol for purification of protein complexes is universal and can be successfully used in different mammalian cell lines. Identified proteins belong to various groups: cytoskeletal, ribosomal, nucleic acid binding, chaperones, enzymes and may potentially be involved in the molecular mechanism of depression.

Affinity Tags for Protein Purification

2020 ◽  
Vol 21 (8) ◽  
pp. 821-830
Author(s):  
Vibhor Mishra
Keyword(s):  
Protein Purification ◽  
Protein Complexes ◽  
Affinity Purification ◽  
Protein Domain ◽  
Affinity Tag ◽  
Small Peptides ◽  
Affinity Tags ◽  
C Terminus ◽  
Solubility Enhancers ◽  
As Solubility

The affinity tags are unique proteins/peptides that are attached at the N- or C-terminus of the recombinant proteins. These tags help in protein purification. Additionally, some affinity tags also serve a dual purpose as solubility enhancers for challenging protein targets. By applying a combinatorial approach, carefully chosen affinity tags designed in tandem have proven to be very successful in the purification of single proteins or multi-protein complexes. In this mini-review, the key features of the most commonly used affinity tags are discussed. The affinity tags have been classified into two significant categories, epitope tags, and protein/domain tags. The epitope tags are generally small peptides with high affinity towards a chromatography resin. The protein/domain tags often perform double duty as solubility enhancers as well as aid in affinity purification. Finally, protease-based affinity tag removal strategies after purification are discussed.

scholarly journals Proximity biotinylation and affinity purification are complementary approaches for the interactome mapping of chromatin-associated protein complexes

2015 ◽  
Vol 118 ◽  
pp. 81-94 ◽  
Author(s):  
Jean-Philippe Lambert ◽  
Monika Tucholska ◽  
Christopher Go ◽  
James D.R. Knight ◽  
Anne-Claude Gingras
Keyword(s):  
Protein Complexes ◽  
Affinity Purification ◽  
Interactome Mapping

scholarly journals Affinity Purification of Chloroplast Translocon Protein Complexes Using the TAP Tag

10.3791/58532 ◽  
2018 ◽  
Author(s):  
Venkatasalam Shanmugabalaji ◽  
Véronique Douet ◽  
Birgit Agne ◽  
Felix Kessler
Keyword(s):  
Protein Complexes ◽  
Affinity Purification

scholarly journals Isolation of Endogenously Assembled RNA-Protein Complexes Using Affinity Purification Based on Streptavidin Aptamer S1

2015 ◽  
Vol 16 (9) ◽  
pp. 22456-22472 ◽  
Author(s):  
Yangchao Dong ◽  
Jing Yang ◽  
Wei Ye ◽  
Yuan Wang ◽  
Chuantao Ye ◽  
...  
Keyword(s):  
Protein Complexes ◽  
Affinity Purification ◽  
Streptavidin Aptamer ◽  
Rna Protein Complexes

An aqueous two-phase system to pre-purify a heterologously produced siderophore

TECHNOLOGY ◽  
2016 ◽  
Vol 04 (03) ◽  
pp. 135-138
Author(s):  
Mahmoud Kamal Ahmadi ◽  
Samar Fawaz ◽  
Blaine A. Pfeifer
Keyword(s):  
Escherichia Coli ◽  
Natural Products ◽  
Cell Growth ◽  
Complex Nature ◽  
Phase System ◽  
Heterologous Production ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Purification System ◽  
Surrogate Host

Natural products span broad activities and applications; however, their access and production are often limited by native cellular sources. As a result, the heterologous production of a siderophore termed yersiniabactin (Ybt) was completed using the surrogate host Escherichia coli. Post-production and purification steps are complicated by the complex nature of most media used for cell growth, prompting the development in this work of an aqueous two-phase pre-purification system capable of rapidly and simply enhancing the concentration of the target Ybt compound.

scholarly journals Leveraging Curation Among Escherichia coli Pathway/Genome Databases Using Ortholog-Based Annotation Propagation

2021 ◽  
Vol 12 ◽  
Author(s):  
Suzanne Paley ◽  
Ingrid M. Keseler ◽  
Markus Krummenacker ◽  
Peter D. Karp
Keyword(s):  
Escherichia Coli ◽  
Protein Complexes ◽  
Limited Resources ◽  
Genome Database ◽  
Single Strain ◽  
Manual Curation ◽  
Genome Databases ◽  
New Knowledge ◽  
K 12 ◽  
New Protein

Updating genome databases to reflect newly published molecular findings for an organism was hard enough when only a single strain of a given organism had been sequenced. With multiple sequenced strains now available for many organisms, the challenge has grown significantly because of the still-limited resources available for the manual curation that corrects errors and captures new knowledge. We have developed a method to automatically propagate multiple types of curated knowledge from genes and proteins in one genome database to their orthologs in uncurated databases for related strains, imposing several quality-control filters to reduce the chances of introducing errors. We have applied this method to propagate information from the highly curated EcoCyc database for Escherichia coli K–12 to databases for 480 other Escherichia coli strains in the BioCyc database collection. The increase in value and utility of the target databases after propagation is considerable. Target databases received updates for an average of 2,535 proteins each. In addition to widespread addition and regularization of gene and protein names, 97% of the target databases were improved by the addition of at least 200 new protein complexes, at least 800 new or updated reaction assignments, and at least 2,400 sets of GO annotations.

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