2a Peptide Motif
2a Peptide Motif - Related Videos
14:28
Peptide-based Identification of Functional Motifs and their Binding Partners
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Cited by 2 •
2013
Techniques to dissect the mechanisms underlying the secretion of HIV-1 Nef in exosomes are described. Specific short peptides derived from Nef and protein transfection were exploited to determine the structure, function, and binding partners of Nef’s Secretion Modification Region. These procedures have general relevance in many mechanistic studies.
04:00
Immunoaffinity Based Extraction of Ubiquitinylated Peptides: A Technique to Selectively Extract Ubiquitin Tagged Peptides from Purified Peptide Fractions
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2023
This video describes a method to extract and purify ubiquitinylated peptides containing remnant di-glycine peptides from a complex peptide mixture. The presented method may help in identifying original ubiquitination sites in the protein.
07:55
Using SCOPE to Identify Potential Regulatory Motifs in Coregulated Genes
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Cited by 3 •
2011
A straight-forward and robust method to identify potential regulatory motifs in co-regulated genes is presented. SCOPE does not require any user parameters and returns motifs that represent excellent candidates for regulatory signals. The identification of such regulatory signals helps to understand the underlying biology.
09:36
Recombinant α- β- and γ-Synucleins Stimulate Protein Phosphatase 2A Catalytic Subunit Activity in Cell Free Assays
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Cited by 7 •
2017
This publication presents a protocol showing how to measure the activity of recombinant protein phosphatase 2A catalytic subunit (PP2Ac) in response to recombinant α-synuclein, β-synuclein, or γ-synuclein proteins using a simple colorimetric assay. We show findings regarding α-synuclein specificity toward PP2Ac in mouse brain.
03:21
Peptide Purification: An RP-HPLC-based Technique to Extract Peptides from Digested Protein Lysates
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2023
In this video, we demonstrate reversed-phase high-performance liquid chromatography to purify peptides from a digested protein lysate mixture. These purified peptides are then lyophilized and stored for downstream applications. Analysis of peptides can help understand the mechanics involved in cellular signaling and cancer.
FAQs
The ribosome progresses through the conserved 2A sequence but fails to form the peptide bond at the Gly-Pro junction. Translation then resumes on the downstream coding sequence, producing a separate protein rather than one continuous fusion product. This interruption is the key molecular event that allows several protein-coding regions to function from one messenger RNA.
A single messenger RNA can link the production of proteins with related experimental roles, such as an antigen, a reporter, and an immune-regulatory factor. This arrangement keeps their coding sequences within one construct and supports analysis of connected infection-related pathways or cellular responses. It can also simplify experimental designs that would otherwise require separate expression systems.
The motif functions at the junction between coding sequences, so its placement determines which protein is translated before the ribosome continues to the next sequence. A design can therefore organize an antigen, reporter, or immune-regulatory factor in a defined order within one construct. This order is important when planning coordinated studies of expression and pathogen biology.
A basic design begins by selecting the proteins needed for the experiment, such as an infection-related antigen, a reporter, or an immune-regulatory factor. Their coding sequences are then arranged in one construct with 2A motifs separating the intended products. The resulting design can be used to examine coordinated protein expression and its relationship to cellular responses.
These designs are useful when a viral vector or vaccine construct needs to express multiple functionally related proteins from one messenger RNA. Linking antigens with reporters or immune-regulatory factors can reduce the need for separate expression arrangements and support coordinated analysis. The approach is therefore relevant to vector development, vaccine studies, and investigations of pathogen-associated immune responses.
2A constructs can help researchers examine whether an antigen, reporter protein, and immune-regulatory factor are produced within the same expression design. That organization supports studies of gene expression, infection-related pathways, pathogen biology, and cellular responses. In practical terms, the method connects protein production from one construct with interpretation of immune or infection-related experimental outcomes.