Misfolded Protein Analysis

the degeneration of brain tissue in mammals. A form of proteins that have undergone alteration form

In UNIT 7 you learned about the basics structure as well as the basic functions of deoxyribonucleic acid. You learned that deoxyribonucleic acid carries an “instruction manuel” to produce proteins that are responsible for passing traits from your parents to you. As you demonstrated in the previous activity, deoxyribonucleic acid is made of four nucleotides base pairs: adenine, thymine, cytosine, and guanine. They are often abbreviated as A, T, C, and G. The uniqueness of deoxyribonucleic acid is not particularly in just these 4 subunits, but how they are arranged. In this activity you will recreate protein synthesis.

The discovery from the Scripps Research Institute in Florida shows promising results in tackling down the cause of Parkinson’s, and their outcomes led to a funding by the National Institutional Disorders and Stroke Research (NINDS). Research staff within the campus discovers that many diseases that relate in twisting a protein from its original structure will result in a cellular death but it isn’t due to the deformed shape. According to the article “Scripps Florida Scientists' 'Mad Cow' Discovery” (2015), one primal cause that leads to Parkinson’s is the lack of “NAD+” which later prohibits the proper energy function of the mitochondria. Researchers further delved into the study to find out this is preventable, by providing the misshaped protein

DLB main neuropathological structures are Lewy bodies, nevertheless it also has the Alzheimer’s disease (AD) pathology of senile plaques and neurofibrillary tangles (Pervin, Edwards & Lippa, 2016). Astonishing up to 80 percent of individuals can show AD pathology (Colom-Cadena et al., 2013). Lewy bodies are located in cell cytoplasm, spherical in shape, eosinophilic, neuronal attachments, with a compressed hyaline centre and clear halo. They are made of unusual shortened and phosphorylated proteins and alpha-synuclein is the main component (Hancock, 2011). The alpha synuclein accumulations, termed Lewy bodies (LB) and Lewy neuritis (LN), disturb brain chemicals resulting in complications with movement, thinking, mood and behaviour. In a fit brain, alpha-synuclein is essential in brain neurons, especially at presynaptic part, where cell

Since the protein does not disintegrate, it amasses into neuritic plaques (The Alzheimer's Project, 2009). The neurofibrillary tangles structure inside neurons and are made out of matched helical fibers, neuropil strings and dystrophic neuritis. Nfts cause the breakdown of the vehicle framework, as tau protein tumbles off and starts to cluster together, framing tangles, irritating the correspondence between neurons, prompting neuronal demise showed through dementia. The neuropathological criteria for an AD analysis are focused around the recurrence of the decrepit plaques and the geography of the neurofibrillary tangles (Goetz, 2007). The cerebrocortical decay prompts the decrease of the mind with twenty percent or more. The

Due to the inability of the brain to replace nerve cells, some brain function is lost. The key question in Alzheimer’s disease is, what causes the neuron degeneration (Johnson, 1989)? The focus for finding the cause is on abnormal structures found in the brain of people with Alzheimer’s. Unfortunately, the abnormal structures the brain undergoes still has researchers uncertain as to how they are involved in Alzheimer’s and exactly how the disease occurs.

Protein purification is a process that can be employed to separate a single protein from a larger starting material which may be anything from an organ to a cell. Isolating a purified protein from a larger fraction enables further analysis such as determination of amino acid sequence, potential biological function, and even evolutionary relationship. (Cuatrecasas 1970) In this experiment, the enzyme lactate dehydrogenase will be purified, this enzyme is found extensively in human cells and catalyzes the conversion of lactate to pyruvate, an essential part in energy production. LDH is a key part of anaerobic energy production especially within glycolysis in which LDH catalyzes the conversion of the reverse reaction, pyruvate to lactate, generating NAD+ from NADH, reproducing the oxidized form of the coenzyme which can be used for oxidative respiration. (Markert 1963) Due to the fact that number of purification steps correlates with the purity of the protein multiple purification techniques will be used to isolate a pure form of LDH. LDH will be isolated from a larger “cytosol” fraction collected from a homogenized rat liver in a previous fractionation exercise. Of the procedures that will be used to isolate and purify proteins from a larger fractionate are a set of techniques collectively known as chromatography. These techniques all have the same premise, in that they consist of a stationary phase, also known as the

Neurodegenerative diseases continue to affect the lives of millions Americans each year, with incidence and prevalence rates ever increasing. These diseases cause degeneration or death of nerve cells in the brain. These diseases can cause a financial and emotional burden on not only patients themselves, but also family members and care givers as well. Molecular mechanisms that underlie these diseases have remained relatively unclear, despite much research. Understanding the mechanisms of these diseases are facilitated by utilizing model organisms to study pathways involved in neurodegenerative diseases. One such model organism is the Caenorhabditis elegans nematode. The C. elegans roundworm has displayed usefulness as a template to study neurodegenerative diseases in humans, including Parkinson’s disease and Alzheimer’s disease.

Purpose: To observe the specificity of protein conformation and to demonstrate the role of enzymes in biological systems that use enzymes; to assess the catalyzing ability of a variety of inorganic and organic molecules and determine the impact of environmental factors on any catalysis occurring in the decomposition of hydrogen peroxide. Hypothesis: When the experiment is performed in an empty vessel there will be no reaction, it is hypothesised this occurred because there is nothing for the hydrogen peroxide to react with. This occurs when sand is in the empty vessel since sand is structurally composed of rocks. Rocks lack functional groups for the substrate to interact with at the active site.

The F-box proteins can be categorized into three major classes based on the presence of specific substrate recognition domains. The FBXW class consists of WD-40 repeat domains that is composed of ten proteins that have need studied extensively. In the FBXL class, there are 22 F-box members that consist of leucine rich repeat protein. The remaining 37 F-box proteins are classified as the FBXO proteins. FBXO proteins are composed of a variety of domains that have yet to be fully analysed and characterized. However, there are promising discoveries attributed to the uncharacterized domains belonging to some FBXO proteins. Greater understanding of these proteins might reveal significant discoveries of how these proteins work and function. Having a large variety of different types of F-box proteins, how do

Alzheimer’s disease or AD is an incurable disorder of the brain that results in loss of normal brain structure and function. In an AD brain, normal brain tissue is slowly replaced by structures called plaques and neurofibrillary tangles. The plaques represent a naturally occurring sticky protein called beta amyloid and in an Alzheimer’s brain, sufferer’s tend to accumulate too much of this protein. Neurofibrillary tangles represent collapsed tau proteins which, in a normal brain along with microtubules, form a skeleton that maintains the shape of the nerve cells. In Alzheimer’s disease, the tau proteins break loose from their normal location and form tangles. Without the support of these molecules, nerve cells collapse and die. As normal

The causes of Alzheimer’s are not yet fully understood; however, its effect on the brain can be understood. Alzheimer’s disease - insidious, attacking and terrifying - stalks and then murders brain cells. A brain afflicted with Alzheimer’s disease has a decreased count in cells and connections among cells. The more brain cells die, the smaller the brain of a person with Alzheimer’s gets. When doctors examine a brain with Alzheimer’s tissue, they see two types of deformities that are known to be trademarks of the disease. The first trademark is known as plaque, which is a cluster of a beta-amyloid protein that may damage and kill brain cells in a number of ways, including blocking with cell-to-cell communication. Whereas the final cause of brain-cell death in Alzheimer’s remains a mystery, the groups of beta-amyloids that cover the brain cells are a sure sign of the disease. The second trademark of Alzheimer’s is the tangle. Brain cells are reliant on internal support and a transport system to bring nutrients and other essentials to their distant regions. This system needs a protein called tau. In Alzheimer’s, strings of tau protein roll into abnormal tangles inside brain cells, concluding in failure of the cell's transport system. This breakdown of the system is powerfully involved in the decline and death of brain

The protein misfolding induces structural conversion of a soluble protein to insoluble amyloids through self-assembling. The protein aggregation induces the loss of biological function and gain of disease and is well connected to several diseases such as neurodegenerative disorders, prion diseases and type-II diabetes. Several studies have been carried out to elucidate the role of protein misfolding and aggregation in the pathogenesis of a number of protein conformational diseases (ref). Among several conformational diseases, the Alzheimer’s disease (AD) has been studied to a wide extent from decades, and a recent study estimated 5.5 million Americans are living with this fatal disease (ref). It also has been estimated (World Alzheimer

Proteins play a part in every cellular activity and must fold into proper three dimensional configurations, or native state, to execute their intended functions. Proteomic stressors such as chemical exposure or elevated temperature can inhibit protein folding upsetting protein homeostasis and resulting in cell death and human disease (Broadley 2009). Excessive protein misfolding can lead to amyloid fibrillar aggregates which deposit around brain neurons contributing to the development or progression of neurodegenerative disorders: Alzheimer’s disease, Huntington’s disease and Parkinson’s disease are all fundamentally diseases of protein misfolding (Soto 2003, Hartl 2009).

Different techniques and principles for protein extraction and characterization were demonstrated in this experiment. Various proteins were extracted from different sources: 1.67 g yeast invertase, 1.03 g egg white albumin, and 5.15 g of milk casein. Activity assay for invertase was performed using Benedict’s test and the enzymes inverting action on sucrose was confirmed. Warburg-Christian Method and Bradford Assay were also employed to determine the protein concentration in the albumin and the casein samples. The concentrations for the albumin and casein samples were found to be 0.519 and 0.327 mg/mL, respectively based on Warburg-Christian Assay; and 6.5x10-3¬ and 1.9x10-2 mg/mL