Infective Prion Analysis

Creutzfeldt-Jakob Disease: It happens when a prion protein misfolds itself, thus causing a “domino effect” which unfolds itself causing a malfunction.

Proteinaceous Infectious Particles, commonly known as Prions, are extremely rare misfolds of the protein PrPc, which cause fatally neurodegenerative diseases, and are theorized to be infectious only by the protein itself (U.S National Library of Medicine, 1998). This “protein-only theory” is still heavily debated today, as some scientists deny the theory, and there isn’t a significant amount of evidence on each side to qualify the theory or disprove it (Soto, C. 2011). The base “Prion” protein is encoded in the gene PRNP, while being non-infectious. Prions are most commonly found in human prion diseases, but they can also be in other animals in the form of Mad Cow Disease and Chronic Wasting Disease, classified as Bovine Spongiform Encephalopathies

Humans have to deal with many different diseases and the ones most disliked are the ones with no cures. Like cancer, transmissible spongiform encephalopathies have no cure, but they are more rare. These diseases are prion diseases which cause the brain to deteriorate. Prions are proteins that sometimes behave like viruses, which mean that they should have some form of nucleic acid, but since they don’t, they cause abnormalities. The nervous system contains many normal prions, but when an abnormal prion comes along, it transforms all the normal prions into abnormal ones. Bovine spongiform encephalopathy is found in cattle, but it can be transmitted to humans.

In bovine spongiform encephalopathy (BSE), the disease is caused by the misfolding of proteins that cause proteins and peptides to develop a fibrillary structure. The PrPc is a correctly folded prion and the misfolded form is called PrPSc. BSE occurs when the normal PrPc come into contact with the toxic PrPSc and the normal prion takes on the shape of the PrPSc. The normal chaperones are unable to convert the PrPSc back to the normal form. The PrPSc now takes on the role of chaperone and the conversion of PrPc prions continue over and over. PrPSc, now being hydrophobic avoids the water of the inner cell and begin to accumulate and form plaques along the neuronal cell membranes. The aggregation of the prions on the cell membrane eventually lead to cell death which produces the sponge-like appearance in the brain of cattle infected with BSE (Thompson, 2014).

Nipah virus, Arena Virus and Francisella tularensis are bioterrorism agents. They work in various ways to harm the host. Francisella tularenis is considered a Tier 1 bioterrorism agent and Nipah virus is an overlap select agent that affects humans and agriculture both. They have been harmful in the past. Though, Nipah virus is a newer virus than the other two. This review will focus on their emergence, pathogenicity and symptoms of the diseases they cause.

Second, in order to further confirm the information about characteristics and function of the targeting protein that we have

4a). This protein had the characteristics of two interactions and a 544 amino acid residue. To further support the claim of the TPP1 interaction with POT1, TPP1 antibody was pulled down with flag tagging POT1 (Fig. 4b). Hence, these data validate the claim of the POT1, and TPP1 form one complex [17].

() The premier levels of this protein are found in the Central Nervous System neurons and the testes. (pro) These proteins are also located in smaller amounts in the liver, lungs, and heart. Localization studies show that the protein appears in the nucleus, endoplasmic reticulum, mitochondria, and Golgi complex. (EJN) This protein has a very distinct portion called the polyQ stretch. (Cattaneo 2005 ) A polyQ stretch is made up of 34 or more glutamine residues. (Cattaneo 2005) This region is found in many transcriptional factors and other disease causing proteins. (Cattaneo 2005) The exact function of this protein is not known, but there is information known stating that it is necessary to have them in early development and nerve cell stability. (Cattaneo 2005) A study was done to show what happens to a mouse embryo by completely disabling the normal Huntington protein. The results of the disruption caused the embryo to die on day 8.5. This occurred before gastrulation and the development of the nervous system. (Cattaneo 2005) This supports the fact that a normal Huntington protein aids in the early stages of development. After the gastrulation process, the Huntington protein becomes necessary for neuron stability. The study for this had less than fifty percent of the wild type Huntington protein in the mice. This revealed a weakness in the neural structure called epiblast. (Cattaneo 2005) The

A prion is a pathogen that lacks nucleic acid. Though they cannot reproduce and are not susceptible to procedures that break down the nucleic acid, they can replicate by stimulating normal cellular prion protein to refold from normal prion protein to abnormal prion protein. They replicate in neural tissue causing degeneration and death. The actual process of how normal prions convert to abnormal prions remains unknown.

The prion protein (PrP) is unusual in that it has two stable isoforms. The cellular or normal form of the prion protein is termed PrPC, while the disease form is termed PrPSC. Sharing the same sequence of amino acids, or primary structure, PrPC and PrPSC differ in their secondary, tertiary, and quaternary structures. The normal prion protein has N- and C-terminals, three alpha helices, and two beta sheets (Soto, 2006, 40). Its function is still not completely understood. However, scientists have found evidence that point to various possibilities. 1) Because most PrPC are located in lipid rafts, membrane structures involved in signaling, it is suggested that PrPC may also be a mediator in neuroprotective signaling pathways. 2) Interaction between PrPC and Bcl-2, a ligand involved in protecting neurons from apoptosis, suggests the possibility that PrPC may be an antiapoptotic protein. 3) PrPC has also been linked to copper metabolism in the brain. In one particular study, PrP knockout mice, or mice that were genetically altered to not express PrP, were found to have lower levels of copper in their brains and higher levels in their serum (Soto, 2006, 42). However, in other studies with PrP knockout mice, most of the animals did not display any obvious disorders, and those that did were found to have been affected by the increase in concentration of another protein known as doppel.

Bettelheim, Brown, Campbell and Farrell assert that polypeptide chains do not extend in straight lines but rather they fold in various ways and give rise to a large number of three-dimensional structures (594). This folding or conformation of amino acids in the localized regions of the polypeptide chains defines the secondary structure of proteins. The main force responsible for the secondary structure is the non-covalent

We created A549 cells with DJ-1 knockout using CRISPR/CAS9 method (Fig…). In this SA, we will determine the effect of DJ-1 deletion on mitochondrial physiology in these cells. It has been reported that mitochondrial dysfunction leads to ROS overproduction (26). We will analyze mitochondrial function by measuring mitochondrial potential (m), mitochondrial energetics, Ca2+ dynamics and ROS generation using confocal microscopy with live cell imaging (Zeiss LSM 510, available at Temple University). A549 cells will be loaded with 50 nM tetramethylrhodamine methyl ester (TMRM) which is a cell permeant dye that

Molecular chaperones stabilize unfolded or misfolded proteins until native conformations have been obtained to promote cell survival during and after stress conditions. They do not change or add to the folding principles encoded by a protein because polypeptide chains inherently carry within them all the information that is necessary for achieving the native state of a protein. Instead, they optimize the folding process by stabilizing folding intermediates and are involved in every aspect of proteome maintenance including de novo folding, refolding of stress-induced misfolded proteins, and targeting proteins for degradation (Hartl 2009, Hartl 2011). Chaperones, many of which are induced or upregulated only during stress conditions, work in cooperative networks when protein-aggregate concentration

Prions are infectious agents composed entirely of a protein material which is transmissible to other prion proteins leading to disease in a manner that is epidemiologically comparable to the spread of viral infection. Prions composed of the prion protein are believed to be the cause of transmissible spongiform encephalopathies among other diseases. All known prion diseases are untreatable and fatal.

Mitochondrial Membrane Potential Detection JC-1 Kit (BDTMMitoScreen) and FITC Annexin-V-Apoptosis Detection Kit I (BD PharmingenTM) were obtained from BD Biosciences. Lipofectamine® 2000, OptiMEM, and JC-1 (5,5’, 6,6’- tetrachloro-1,1’,3,3’tetraethylbenzimidazolocarbocyanine iodide) were purchased from Life Technologies and Molecular Probes respectively. TUNEL assay kit, Dual luciferase reporter gene assay kit and Proteasome-GloTM assay reagents were obtained from Promega. Anti-ubiquitin, anti-GFP, anti-20S proteasome, anti-p53, anti-p27, anti-p21, anti-Bax, anti-IkB-α, anti-β-actin and anti-HA antibodies were purchased from Santa Cruz Biotechnology. Horseradish peroxidase-conjugated anti-rabbit-IgG and anti-mouse IgG; anti-rabbit, and anti-mouse (IgG fluorescein isothiocyanate and IgG-rhodamine) were obtained from Vector Laboratories. Plasmid pcDNA™ 3.1 was purchased from Life Technologies. pcDNA3-EGFP (Addgene 13031), pRK5-HA-Ubiquitin-WT (Addgene plasmid 17608), pEGFP-Hsp70 (Addgene plasmid 15215) and Luciferase-pcDNA3 (Addgene plasmid 18964) plasmids were purchased from Addgene.