Alpha Factor Secretion Signal
Alpha Factor Secretion Signal - Related Videos
09:03
Monitoring Functionality and Morphology of Vasculature Recruited by Factors Secreted by Fast-growing Tumor-generating Cells
0 Views •
Cited by 7 •
2014
We describe a matrigel plug assay to illustrate angiogenic potential of a pool of factors secreted by cancer cells using two complementary imaging modalities, ultrasound and endomicroscopy. The matrigel, an extracellular matrix (ECM)-mimic gel, is utilized to introduce the host (mouse) with angiogenic factors secreted to the conditioned media...
11:33
Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
0 Views •
Cited by 2 •
2013
This article illustrates the use of pulse-chase radio labeling in combination with site-specific photocrosslinking to monitor interactions between a protein of interest and other factors in E. coli. Unlike traditional chemical cross-linking methods, this approach generates high resolution “snapshots” of an ordered assembly pathway in a living cell.
04:08
An Assay for TLR-Dependent NF-кB/AP-1 Transcription Factor Signaling in Macrophages
0 Views •
2025
This video demonstrates an assay to examine the toll-like receptor(TLR)-dependent activity of the transcription factors nuclear factor-kappa-B (NF-κB) and activator protein-1 (AP-1) in macrophages. The TLRs trigger a pro-inflammatory signaling cascade upon recognizing damage-associated molecular patterns (DAMPs), activating NF-κB and AP-1. The engineered reporter macrophages produce secreted embryonic alkaline phosphatase (SEAP) enzyme upon NF-κB and AP-1 activation, which is detected via a...
03:11
Quantification of Bioactive Type-I Interferons using a Secreted Embryonic Alkaline Phosphatase Reporter Assay
0 Views •
2025
This video demonstrates secreted embryonic alkaline phosphatase assay using HEK-Blue interferon-α/β reporter cells to quantify the type-I interferons (IFNs) released following stimulation of plasmacytoid dendritic cells by HIV-1 Infected CD4+ T cells.
08:51
Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling
0 Views •
Cited by 11 •
2015
The method described here is used to induce the apoptotic signaling cascade at defined steps in order to dissect the activity of an anti-apoptotic bacterial effector protein. This method can also be used for inducible expression of pro-apoptotic or toxic proteins, or for dissecting interference with other signaling pathways.
FAQs
The prepro-leader operates as a sequence of linked stages rather than as a single permanent tag. It first guides the fusion protein into the endoplasmic reticulum, where the signal peptide is removed. Subsequent proteolytic processing then prepares the secreted product for release, connecting intracellular trafficking with maturation of the protein.
Kex2 and Ste13 contribute at the processing stage of the pathway. Their activity acts on the alpha-factor-derived leader so that the attached recombinant protein can be released as a mature product. Examining this processing is therefore useful when interpreting whether poor recovery reflects trafficking, incomplete maturation, or secretion efficiency.
Successful secretion depends on more than delivery to the endoplasmic reticulum. The protein must also undergo appropriate proteolytic processing and folding, and the overall trafficking pathway must function effectively. These linked requirements make secretion output informative: a change in recovered product can reveal limitations in processing, folding, trafficking, or other efficiency-related factors.
Compared with approaches that require cell disruption, alpha factor-mediated secretion places the recombinant product in the culture medium. This changes the recovery problem from extracting protein from cellular material to collecting the external fraction. The result can simplify downstream recovery and reduce dependence on disruption, which is valuable when secretion is efficient.
An experimental workflow can begin by fusing the protein of interest to the alpha factor prepro-leader and expressing that construct in Saccharomyces cerevisiae. Researchers then examine the culture medium for the secreted product and consider whether its processing produced the intended mature form. This workflow links construct design, cellular trafficking, extracellular recovery, and product assessment.
In biotechnology, the signal supports production of recombinant proteins without making intracellular extraction the central recovery step. In biology, it also provides a system for studying how eukaryotic cells traffic, process, and fold proteins during secretion. Measuring the resulting product in culture media can therefore address both engineering goals and questions about secretory pathway performance.