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- Table of Contents
Facts about Apoptosis-associated speck-like protein containing a CARD.
Involved in activation of the mitochondrial apoptotic pathway, promotes caspase-8-dependent proteolytic maturation of BID independently of FADD in certain cell types and also mediates mitochondrial translocation of BAX and triggers BAX-dependent apoptosis coupled to activation of caspase-9, -2 and -3. Involved in macrophage pyroptosis, a caspase-1-dependent inflammatory type of cell death and is the primary constituent of the ASC pyroptosome which forms upon potassium depletion and rapidly recruits and activates caspase-1.
Human | |
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Gene Name: | PYCARD |
Uniprot: | Q9ULZ3 |
Entrez: | 29108 |
Belongs to: |
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No superfamily |
ASC; ASCMGC10332; CARD5; CARD5TMS; caspase recruitment domain protein 5; Caspase recruitment domain-containing protein 5; hASC; PYCARD; PYD and CARD domain containing; PYD and CARD domain-containing protein; Target of methylation-induced silencing 1; TMS1; TMS-1; TMS1apoptosis-associated speck-like protein containing a CARD
Mass (kDA):
21.627 kDA
Human | |
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Location: | 16p11.2 |
Sequence: | 16; NC_000016.10 (31201486..31202776, complement) |
Widely expressed at low levels. Detected in peripheral blood leukocytes, lung, small intestine, spleen, thymus, colon and at lower levels in placenta, liver and kidney. Very low expression in skeletal muscle, heart and brain. Detected in the leukemia cell lines HL-60 and U-937, but not in Jurkat T- cell lymphoma and Daudi Burkitt's lymphoma. Detected in the melanoma cell line WM35, but not in WM793. Not detected in HeLa cervical carcinoma cells and MOLT-4 lymphocytic leukemia cells.
Cytoplasm. Endoplasmic reticulum. Mitochondrion. Nucleus. Upstream of caspase activation, a redistribution from the cytoplasm to the aggregates occurs. These appear as hollow, perinuclear spherical, ball-like structures. Upon NLRP3 inflammasome activation redistributes to the perinuclear space localizing to endoplasmic reticulum and mitochondria. Localized primarily to the nucleus in resting monocytes/macrophages and rapidly redistributed to the cytoplasm upon pathogen infection. Localized to large cytoplasmic aggregate appearing as a speck containing AIM2, PYCARD, CASP8 and bacterial DNA afte
In this article, we'll examine the best way to use the PYCARD Marker in the quantitation of proteins and genes. To conduct quantitative PCR, we will use primers IL-1b or NLRP3. These primers were designed specifically for the PYCARD Marker. The primers are available on Boster Bio's website. First, you must download the appropriate software in order to use the PYCARD Marker.
The PYCARD marker is a highly versatile molecular probe for biomedical applications. This biosensor is capable of reading absorbance at 450 nm and detects the presence of PYCARD v1 protein. It is also compatible with a wide range of cells that include human cells as well as tissues. For the best results, use the PYCARD immunoassay kit.
Amplification efficiency can be measured by quantifying the number of amplification cycle, however, various factors can impact the overall amplification efficiency. Therefore, many applications use an internal standard gene to conduct their PCR reactions. This housekeeping gene is a second primer pair used to verify the accuracy of the target. Quantitative PCR costs less than basic PCR and requires less cycles to amplify the target.
Quantitative PCR primers must be specific to a particular gene. To ensure that the primers generate an exact product they must be derived from known reference genes that are stably expressed. For a more stable cDNA yield choose primers that have threshold cycle (Ct) values within the range of mean +-1 for each reference gene. It is recommended to employ two sets to test the quality of the cDNA that is generated.
RT-qPCR (also known as Quantitative Real-Time PCR) is a combination of amplification and quantification of the target DNA sequence. The resulting product contains a variety of fluorescent labels and allows for quantifying copy numbers. Additionally, qPCR has also been developed for numerous applications including detecting pathogens and measuring the number of copies of DNA sequences that are of interest. It is the standard method for verification of microarray gene expression data.
PCR is a common laboratory technique. It involves using an enzyme called DNA polymerase to create multiple copies of a particular DNA region. The area being copied can be anything, from a gene function to a genetic marker. Forensic scientists are able to use PCR to match DNA from crime scenes to a suspect. The amplified DNA could be sequenced and examined using gel electrophoresis. It is also possible to be cloned into viruses for further experiments.
Inflammasomes are multiprotein complexes that contain caspase 1, 5, and 8 and NLRP1 or NLRP3 as upstream effectors. PYCARD, NLRP1, and IL-1b are all upregulated in AD. Interestingly, only a small percentage of MCI patients develop AD, suggesting that PYCARD and IL-1b are prodromal markers of the disease.
CAPS is caused by IL-1b. MCC950, an inhibitor of MCC950 effectively blocks the activation of NLRP3 by mice, which increases the survival rate and weight. Studies have demonstrated that neutrophils are the main driver of CAPS pathology. This may also include IL-1b. Deficient mice show reduced CAPS pathogenesis when they have gasdermin D. This is in support of the search for selective gasdermin D inhibitors that treat CAPS.
It isn't clear which protein is responsible the activation of IL-1b, but it is implicated in NF-kappaB activation. Inflammatory mediators like IL-18, IL-1b and have been implicated in inflammatory responses. Using IL-1b and IL-18 as a PYCARD marker can help researchers understand how the two components of the inflammasome work.
Pycard genes encode a protein that looks like a speck, the inflammasome adaptor proteins. This protein interacts to the sensor protein and creates tiny specks within the cytosol. Once activated, active Caspase-1 dimers join with the speck. Pro-IL-1b and IL-18 are then activated, and then cleaved into cytokines that are biologically active.
The PD pathophysiology is critical because of the pyroptosis reaction in the inflammation response. Caspase-1 is activated by a multiprotein signaling complex called the inflammasome, and NLRP3 is a major regulator of the inflammasome. In in vitro, inhibition of NLRP3 by MCC950 significantly decreased the expression of inflammatory cytokines caused by PD in C2C12 cells.
PYCARD is an epitope found on human leukocytes and plasma cells. It is linked to various inflammation processes, including the induction of chemokines or cytokines. Chemokines, cytokines are believed to be responsible for the adaptive immune response. In particular, they are involved in muscle immunity. They also play an important role in the transition between monocyte and neutrophil recruitment. They also play an important part in the control of local inflammation in vivo as well as assist in tissue repair. They are involved in the regulation of immune function and IL-1b's inflammation.
PMID: 10567338 by Masumoto J., et al. ASC, a novel 22-kDa protein, aggregates during apoptosis of human promyelocytic leukemia HL-60 cells.
PMID: 11103776 by Conway K.E., et al. TMS1, a novel proapoptotic caspase recruitment domain protein, is a target of methylation-induced gene silencing in human breast cancers.