F-Box Protein Analysis

The purpose of this lab was to purify and test a GFP protein via several laboratory methods for the purpose of purifying and testing the protein in SDS-PAGE. To purify the protein chromatography and gel electrophoresis were the methods used in the experiment. GFP in the samples were tested using an ultraviolet light. When GFP was found present the cell were transformed into a petri dish containing ampicillin and arabinose. The cells were then lysed and SDS-PAGE was used to test.

Before the Market Revolution, America as a whole differed greatly from the America that had developed during the start of the 19th century. The ideals and values of an American family life varied from those after the revolution. American economy had relied heavily on agriculture and people had a more personal work sphere. The government as well, was cautious when involving itself in the nation’s economic affairs. The Market Revolution overall played a big role in changing American society, economy, and politics by fueling sectionalism, increasing commercialism, and forcing the government to take on more responsibilities to benefit the nation.

It can mutate in many ways, but truncating mutation types are associated with the greatest incidence of breast cancer because since DNA repair functions are not functioning properly, other mutations of fast-replicating cells go unchecked and allow for aggressive tumor growth. BRCA2 is the next type of tumor suppressor gene, located on chromosome 12q12 and is also involved with DNA repair, and also result from truncated proteins. BRCA2 mutations account for 35-40% of all BRCA gene mutations, and 28-32% of all hereditary breast cancers. So, what do breast cancers actually do? For starters, we have cytokines that mediate intracellular communication locally to cause an integrated response to stimuli. These can be synthesized be healthy or diseased cells, and their job is to regulate survival, growth, differentiation and effector functions of the cell. In breast cancer, many cytokines have a role in expression of cancer cells and are usually produced at the site of the tumor. The certain suppressive cytokines may inhibit the immune response against tumor cells and will inhibit the functions of lymphocytes that are associated with tumor suppression and may in turn actually promote tumor growth. There is a category of cytokines that are

One study by Shuijun Zhanga et al had shown the ABT-737 induced apoptosis of hepatocellular carcinoma cells (HCC) by the transcriptional repression of Myeloid cell leukemia 1 (Mcl-1). Shuijun Zhanga et a. had shown that NCTD could affect apoptosis by the modification of the B-cell lymphoma 2

Therefore, dose and duration of treatment is limited, which in turn limits the amount of normal and tumor cell death. A second mechanism is the suppression of cancer cells for variable periods of time without cell death. This mechanism is referred to as remission. Unfortunately, the cancer can return at any time, and it is sometimes stronger. Here another limitation is introduced. Some tumor cells can develop resistance to a particular chemical agent, or several chemical agents, limiting the types of chemotherapeutic agents available for effective use. The last mechanism is cell differentiation, which helps the immune system learn to recognize and fight tumor cells (1).

Fig. 3.Immunization of balb/c mice against fibrosarcoma tumor, WEHI-164. Mice were inoculated with 10 μg of each component mentioned above at weekly intervals for three consecutive weeks. Seven days after last vaccination, approximately 2×106 cells/ml of WEHI-164 was injected andthe inhibition of tumor growth was noted every seven days for three weeks in a

MYC oncoprotein is a transcriptional factor that activates genes involved in cell proliferation and growth. Thus translocations or other gene rearrangements involving MYC may promote cell proliferation and is classically associated with high-grade B-cell lymphomas such as Burkitt lymphoma. MYC rearrangements however, are not unique to Burkitt lymphoma and have been observed in other B-cell lymphoproliferative disorders, including subsets of diffuse large B-cell lymphoma (DLBCL), transformed follicular lymphoma, blastoid mantle cell lymphoma, plasma cell myeloma, and lymphoblastic lymphoma. Aberrations in MYC have been reported rarely in de novo B-cell prolymphocytic leukemia (B-PLL). In CLL, aberrations of MYC are a very rare (<1 % of CLL)

For the second part of the experiment, one had to use the knowledge learn from viewing protein molecules in FirstGlance in Jmol to analyze the protein PDB ID: 4EEY. The analysis of this protein was done using the RSCB protein data bank (PDB) at (http://www.rcsb.org/pdb/home/home.do).2

Previous studies have suggested that K63 ubiquitination is associated with anti-apoptotic protein trafficking pathways. Figure 1 addresses whether K63 ubiquitination is accomplished by the TRAF6 E3 ligase. An immunoblot of p53 was used for U2SO cells in a Ni-NTA assay to determine if K63-linked ubiquitination of p53 was associated with TRAF6. U2SO cells were used because of their high proliferation rates. Flag-TRAF6 and p53 were included in the western blot as controls for this assay. Ubiquitin was tagged with histidine and TRAF6 was tagged with the Flag antibody to assess protein to protein interactions. This assay shows ubiquitination of p53 in the presence of His-Ub and Flag-TRAF6 indicating that K63 linked ubiquitination of p53 depends on TRAF6. To determine if specifically, K63-linked ubiquitination occurs through TRAF6 the authors assessed whether TRAF6 promotes K48-linked ubiquitination of p53 and proteasomal degradation. An immunoblot for HA in a Ni-NTA pull-down assay was generated to assess if ubiquitination and proteasomal degradation occurs at K48 and K63 by TRAF6. HA-p53 and Flag-TRAF6 in U2SO cells were used to show protein-protein interaction. A

This begs the question upon how the DEK protein is able accomplish this task, by evaluating the biological pathways that DEK utilizes. The most commonly inactivated tumor suppressor gene, is the human p53 protein. Despite this, the protein has a short half-life and undergoes continual proteolytic degradation in healthy cells, regulated by the E3 ligase MDM2 [31]. MDM2 works by physically up taking the trans activation domain of the p53 and will thereby prevent its transcriptional production [32]. However, the feedback loop of the MDM2- p53 complex can be disturbed by stress due to DNA damage or lack of oxygen leading to cell death [30]. Bax, is a cell death inducing factor that belongs to the Bcl-2 family. Under regular conditions it forms in the cytosol and adheres to the membrane. Once the cell has been signaled for apoptosis, Bax will translocate into the mitochondria where it will become an integral protein of the membrane. Cross-linking Bax proteins will initiate for the expulsion of apoptotic factors from the mitochondria in order to initiate the apoptotic pathway [35-38]. Next, these

Even though BCC is genetically different from other types of cancers due to its inability to metastasize, the identification of these new drivers can offer a starting point for other researchers to investigate the potential drivers in other forms of cancer that are more complex and have genomic instability. In some ways, BCC has already begun being compared to cancers such as colorectal cancer and pancreatic cancer. Preclinical studies demonstrated the antitumor activity of vismodegib in xenograph models of colorectal and pancreatic cancer. Similarly, phase 1 clinical trials in advanced BCC highlighted an objective response to vismodegib (De Smaele et al.). Due to vismodegib’s low toxicity and specificity for the Hh pathway, this drug has potential advantages compared with conventional chemotherapy, and may eventually be used in combination

Secretion of the protein can either be to become a part of the plasma membrane as an integral protein or free-floating in the cytoplasm so it can be used within the cell such as the protein coded by eif-3.F. The protein encoded by eif-3.F plays a role mainly during the initiation phase of eukaryotic

Cancer may be an organic {process|biological method} process Cancer is a multi-step process that is driven by genetic mutations and epigenetic alterations of deoxyribonucleic acid. The genetic and epigenetic alterations generally have an effect on the genes dominant growth, proliferation and survival of the cells. It’s long been recognized that cancer is associate degree organic {process|biological process} process that is characterized by accumulation of mutations that drive tumour initiation, progression and development of treatment resistance. This advanced method needs bypassing a series of barriers between traditional and cancer cells and reflects the work of evolution. Associate degree early theory

Normally in healthy cells, the protein kinases (PK) may act as proto-oncogenes or tumour suppressor. However, alteration in PKs may lead to development of cancer by several mechanisms, including the activation of cell multiply pathways, genomic instability, diminish of apoptotic pathways, the endorsement of angiogenesis and cell motility. Receptor and non receptor TKs are mostly deregulated in several types of cancer. EGFR is a transmembrane receptor kinase that is overexpressed or aberrantly activated in NSLC.

The body is affected on a cellular level by cancer. Going more into detail, cancer is caused by an error in the replication process of a cell. The cell is changed to rapidly and relentlessly reproduce more replicas of its’ mutated self. These cancer cells, along with the power to multiply quickly, have the innate ability to ignore commands from the body about destruction. If there are ever too many cells in the body, messages are sent to perform apoptosis. Apoptosis is where the cell commits suicide, but cancer cells can become immune to this signal. Together the two combine to make an unstoppable cell growing factory.