Cell homeostasis is a complex yet strictly regulated by many regulators. Different from big role-player such as enzymes in many signaling pathways, the universal consensus of small regulator ubiquitin contributes a great length to the of the complex post translational regulating protein system. Consisting of 76-residue protein, ubiquitin not only highly expresses in cell for various signal transduction pathways, it also is one of the most resilience and packed protein that is highly conserved among the eukaryotes[1]. Ubiquitin is firstly produced intracellularly as precursor fusion protein chain consisting polyubiquitin or ubiquitin fused to amino-terminus of Ribosomal protein 40S and 60S units; the generation of free ubiquitin from these precursor …show more content…
The de-ubiquitinase (DUB) will cleave this precursor into free monomer ubiquitin molecules and can be activated and attached to the target protein through ATP dependent process called ubiquitination with the help of activating enzyme E1, conjugating enzyme E2, and ligase E3. Through forming (iso)-peptide, thioester, or ester bond between carboxyl groups of C-terminal glycine 76 on ubiquitin and the ε-amino group of the lysine, cysteine, serine/threonine on the substrate, respectively, the poly-ubiquitin chain is formed [3]. Based on the different internal Lysine (Lys6, Lys11, Lys27, Lys29, Lys33, Lys48, and Lys63) and on the proximal ubiquitin, the (iso)-peptide bond can be form with another ubiquitin, distal mono-ubiquitin [4, 5]. Consequently, seven conventional poly-ubiquitin chains resulting in different 3-dimentional shapes explaining its different characteristics inherited upon binding to the target protein [5, 6]. With ranging more than 500 putative E3 ligases exist in human, the E3 ligase will tag the poly-ubiquitin chain to the target protein with high specificity
The purpose of this lab was to identify unknown bacteria cultures using various differential tests, and my unknown bacteria is #17. The identification of these unknown cultures was accomplished by separating and differentiating possible bacteria based on specific biochemical characteristics. Whether the tests performed identified specific enzymatic reactions or metabolic pathways, each was used in a way to help recognize those specifics and identify the unknown cultures. The differential tests used to identify the unknown cultures were Gram stain, Catalase, Mannitol Salt Agar (MSA), Blood Agar, Novobiocin, Coagulase, and DNAse (Alachi, 2007).
A) Describe in your own words, in as much detail as you can, the anaerobic metabolism of glucose to pyruvate. B) Draw this pathway (by hand), indicating all substrates, enzymes, cofactors and products. (You do not need to include reaction mechanisms.)
Measure the initial width, length, and thickness of the steel specimen using a Dial Caliper. Relieve pressure in Amatrol T9014 and adjust the height of the bottom platform to insert steel specimen. Insert one pin into the bottom platform to hold the steel specimen into the fixture. Slide two locking bars down the steel specimen. Adhere one locking bar to the bottom of the specimen and one at the top, lock them in place using the attached thumb screws. Insert the Linear Vernier Caliper in the top locking bar and zero out the caliper, allowing it to rest on the bottom locking bar. Compress the hydraulic cylinder until the indicator reads a force of zero. Lock the Linear Vernier Caliper in place by tightening the top thumb screw. [1] Compress
Imatinib is a Abl/c-kit/PDGFR inhibitor. Abl is a proto-oncogene related with chronic myelogenous leukemia. c-kit is a protein on the surface of various types of cells. PDGFR is a cell surface tyrosine kinase receptor for members of the platelet-derived growth factor family.
There are many types of ubiquitin ligases in cells. However, we can make some candidate groups of targeting E3 based on the bioinformatics database.
The two independent variables were luminant cue patches (light cue, dark cue and equiluminant cue) and location of the cue and target (valid side with cue and target on same side and invalid side with cue and target on opposite sides). The dependent variable was participants’ reaction time in millisecond.
The hospitals are large and serve many patients, so there is a lot of lab works needed. Each hospital is different in running samples and type of instruments used. The first hospital we visit was UAB hospital. The UAB hospital has a big lab and many samples are analyzed. The lab is fully automated, all the sample have a barcode and the samples directed to the instruments depend on what test need to be performed. There are 6 millions different test running a year. They spend around 25000 $ on reagents and 15000 on controls every year.
Pre-mRNA in eukaryotes is spliced by the spliceosome, an RNA-protein complex, in which U1, U2, U4, U5 and U6 snRNPs are equal components. However, U1 is more abundant than other snRNPs in cells. U1 snRNP has been found to functions other than splicing, namely in protecting pre-mRNA from premature cleavage and polyadenylation. This protective role may account for its increased levels within cells.
Sample preparation: purified protein and cell lysate have been already and denatured in Laemmli sample buffer and are in His-tagged DHFR, GST-tagged DHFR, Myc-Flag-tagged DHFR aliquots. Control lysate is provided by TA. Wear goggles before to hreat samples for 2 minutes at 95˚C. Centrifuge samples and load gel at room temperature. Obtain an aliquot of the each of the following: 1X Laemmli buffer and Kaleidoscope prestained protein standard (Bio-Rad
The purpose of these series of experiment was successful as seen from the results. The recombinant form of the green fluorescent protein was successfully expressed and purified from E. coli. Though a high yield and purity were not obtained, the qualitative monitoring of the rGFP activity was consistent with the quantitative values obtained. Qualitatively, the elution fraction E2 fluoresced the most when placed under a hand-held UV lamp. Quantitatively, E2 also had the highest activity with 41900 RFUs when read by a fluorescent microplate reader. One way to increase the yield of rGFP would be to increase the IPTG induction time. This would increase the amount of rGFP that is expressed and consequently increase the amount of rGFP that is purified. There were other things during the experiment such as contaminant proteins that could also have affected the yield of rGFP obtained.
The central domain (residues 34-269), containing eight twisted β-sheet strands with coinciding α-helices, holds DNA and ATP binding sites and is structurally similar to ATPase domains in DNA and RNA helicases. ATP will bind at the phosphate-binding loops of this domain, found in many proteins involved in ATP binding (7). Amino acids lysine and threonine live near this P-loop and directly interact with phosphate groups in ATP. Alongside, two disordered loops named L1 and L2 bind to ssDNA and the duplex DNA. What is important about L1 and L2 is that because they support one of the essential functions for RecA in DNA binding, they are highly conserved. Between the α -helix G and L2 boundary lie two glycine residues that give a greater interaction between the positively charged amine groups of DNA helix and the negatively charged sugar-phosphate backbone in DNA molecules
The Miller-Urey Experiment was conducted in 1952 to test if a primitive Earth environment were reconstructed with the necessary amount of energy, then the formation of some organic complex molecules (amino acids) should form. Since our planet is billions of years old, our atmosphere has change significantly since they introduction of life. Since amino acids are the building blocks of all life we know of, the explanation of how they came to be is important. This experiment is fundamental when discussing the origins of life on Earth and how plausible the formation of organic molecules was in these early conditions.
Ultraviolet light will affect the growth of basil. The plant setup with UV light will grow faster than the setup without UV-B light. To determine whether the experiment is successful I will measure the speed of growth, the number of seeds that germinate after a certain amount of days. Another important factor to consider prior to the laboratory experiment is whether the plant is sensitive to UV-B light or not. Sensitivity is most noticeable with regard to alterations in plant growth and development (Teramura 1983). UV-B radiation can damage the DNA structure of the plant and generate growth issues to the cells of Ocimum basilicum. However, the perception of low levels of UV-B by plants might actively promote survival because it may
Birinapant (TL32711) is a bivalent SMAC mimetic. Apoptosis resistance acquisition is a fundamental event in the development of cancer. Among the mechanisms is the dysregulation of inhibitor of apoptosis (IAP) proteins regulated by endogenous IAP antagonists such as SMAC. Drugs mimicing the SMAC have been designed to overcome IAP-mediated apoptosis resistance of cancer cells.
The starting materials, CA and 2-IZT were obtained from Sigma-Aldrich (Mexico) and used as received with ultrapure Millipore water in all experiments. In a typical preparation procedure, CA (101 mg) and 2-IZT (161 mg) were dissolved in water, and then sonicated and vigorously stirred to make the solute completely dispersed with the use of an Ace pressure tube flask from Sigma-Aldrich, followed by vaporization at 70 ℃ in magnetic stirrer until dry within 12 hours (h). Additionally, 10 ml of ultrapure water was added to the resulted dense syrup, and then the solution was heated at 180 ℃ and stirred for 24 h. After the reaction completed, the flask was cooled to room temperature. The obtained final product appearance was a transparent solution