Study of the folding and unfolding of proteins adsorbed to a fused silica surface
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This document summarizes a study on the folding and unfolding of cytochrome c proteins adsorbed to a fused silica surface. It introduces cytochrome c, describes solution and surface spectroscopy experiments, and compares experiments using horse and yeast cytochrome c. Specifically, it examines how horse cytochrome c unfolds with increasing alcohol concentration in solution versus on the surface, finding proteins are more protected from denaturation when adsorbed. Similar experiments with yeast cytochrome c show it takes longer to unfold on the surface than in solution.
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The document summarizes research on the adsorption of cytochrome c (cyt c) proteins to a fused silica surface at varying concentrations. The following key points are made:
1. Using ATR spectroscopy, the researchers found that cyt c proteins adsorbed to the surface at lower concentrations were more denatured than those at higher concentrations.
2. As the surface coverage of cyt c increased with higher concentrations, protein-protein interactions led to greater renaturation of the adsorbed proteins.
3. The degree of renaturation depended on the surface coverage, with maximal renaturation observed at saturating concentrations above 15-20 μM, when protein-protein interactions were highest.
Phyto-Course-Brodbelt-2013.ppt
This document discusses analytical chemistry methods for the identification and quantitation of compounds in complex mixtures. It focuses on the analysis of flavonoids using mass spectrometry and chromatography. Key points covered include sample preparation techniques, spectroscopic methods like UV-Vis, IR, NMR and mass spectrometry for structural analysis, and calibration curves for quantitative analysis. As an example, the document describes the specific analytical workflow used to analyze and quantify the flavonoids quercetin and kaempferol in kale extracts using LC-MS/MS.
Modification & Application of Borate Zirconia Catalyst
Solid catalysts are of great advantages in alkylation reaction due to heterogenous reaction which makes separation of catalysts very easy and environment friendly. Here, sulfated and borate zirconia catalysts are used to search for ortho-xylene with Toluene & methanol. To find a new path to get o-xylene, catalysts surface was studied and a new mesoporous borate zirconia catalyst was prepared. Mesoporous Borate Zirconia had showed a very efficient path to manufature o-xylene.
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M1-Biophysics
Following is my journal documentation during Master's in Biotechnology completed in 2015. I do understand many changes would've occurred in the curriculum since then, but the basics seldom change. Kindly absorb as per your need.
P i trap
Microfluidic devices can be used for sample preparation and analysis in proteomics. Pipettes and channels in microfluidic devices mimic traditional lab tools at a smaller scale. Membranes in microfluidic devices allow controlled addition of reagents to samples. Diffusion is faster at the microscale enabling rapid reactions. Electrocapture using microfluidic membranes can isolate and concentrate proteins for downstream analysis like mass spectrometry. PI-trap microfluidic devices use pH gradients created by ampholytes to fractionate proteins based on isoelectric point, improving proteome coverage. These microfluidic approaches show potential for clinical proteomics applications.
C-terminal Sequencing of Protein : Novel Partial Acid Hydrolysis & Analysis b...
The document describes a novel method for C-terminal sequencing of proteins using partial acid hydrolysis and mass spectrometry analysis. Peptides or proteins are hydrolyzed with vapors of strong organic acids like trifluoroacetic acid or hepta-fluorobutyric acid at high temperatures. This results in successive degradation of the C-terminal residues as seen by mass spectrometry. The degradation is believed to occur via formation of an oxazolone ring at the C-terminal amino acid, followed by removal of the C-terminal residue. Specific cleavages also occur at the peptide bonds preceding aspartic acid and serine residues. This method allows efficient C-terminal sequencing of proteins in small quantities directly from mass
Essay On Down Membrane
1. Isopentyl acetate was synthesized via an acid-catalyzed Fischer esterification of isopentyl alcohol and acetic acid. Sulfuric acid was used as the catalyst.
2. The reaction products were extracted with ether and the ether layer was dried over sodium sulfate. Distillation was then used to purify the product.
3. IR spectroscopy, gas chromatography, and comparison to an authentic isopentyl acetate sample were used to identify the final product as isopentyl acetate. The IR spectrum and GC results matched those of the
Hydrophobic interaction chromatography (1)
Hydrophobic interaction chromatography (HIC) is used to purify biomolecules like proteins by exploiting differences in hydrophobicity. It works by separating substances based on their varying strength of interaction with hydrophobic groups attached to an uncharged gel matrix. HIC is useful for downstream purification as it can handle large sample volumes with high salt concentrations and is gentler than other methods, maintaining protein activity. Factors like ligand type, salt concentration, pH, and temperature affect HIC separation. The document evaluates HIC for monitoring plasmid DNA purification, finding it is a simple, rapid and reproducible method to assess purity.
141 Novel Poster final
This document summarizes an investigation into the effects of palladium nanoparticles (PdNPs), pH, and temperature on the reduction of hexavalent chromium (CrVI) to its less toxic form CrIII. Key findings include:
1) The optimal conditions for the reduction reaction were pH 4 and 45°C temperature.
2) Reaction kinetics followed pseudo-first order behavior, indicating a linear decrease in CrVI over time.
3) PdNPs catalyzed the reaction, decreasing the activation energy by 20±10 kJ/mol and increasing the rate constant over an order of magnitude.
4) Under optimal conditions, a 26% reduction of CrVI was achieved.
CHM-501-SERMACS poster final draft
This document describes a study that aimed to develop a method to determine isoflavones in the sea anemone Aiptasia pallida using solid phase extraction (SPE) and gas chromatography-mass spectrometry (GC-MS). SPE was used to isolate analytes from symbiotic and aposymbiotic anemone seawater. GC-MS analysis with different solvents like methanol, hexane, and dichloromethane showed minor peaks in seawater samples but results were inconclusive. Derivatization of samples with BSTFA prior to GC-MS produced distinguishable peaks, but the method needs further optimization to better separate peaks and identify specific isoflavones. More experimentation is also needed to
Acido urico
This paper studies the kinetics of uric acid dissolution at different pH values using turbidimetric measurements. The rate of dissolution is proportional to the difference between the uric acid concentration at equilibrium and the current concentration, indicating it follows first-order kinetics. Experiments using three different uric acid seeds show the dissolution rate constant decreases as pH increases. Analysis of the results supports the dissolution process being controlled by the aqueous boundary layer, as is typical for organic compound dissolution.
Friccohesity and survismeter: New state of physicochemical science GU 22 Nov ...
Shear stress and surface energy are the optimizing data to truly conduct the activities in the cell. A high or low shears initiate a remarkable ability to induce exponential activities in biochemical processes. Increasing shear generates a big in chemical potential of high shear and low shear liquids so a chemical system induce osmotic pressure. The high shear liquids attract low solvent of a shear liquid. Since there is a mass transfer by osmotic pressure where electric or electrostatic potential influence the mass transfer. Thus, the conductance is naturally involved in the process. Lecture covers the fundamental sciences of the friccohesity and survismeter and their potential to connect with shear stress, surface energy, osmotic potential, conductance.
Spectrophotometry lecture
This document discusses spectrophotometry techniques for measuring light absorption by molecules. It covers the electromagnetic spectrum, Beer-Lambert law, applications of UV-Vis spectroscopy like determining cell density and protein concentration, and methods for protein concentration measurements including Bradford assay and measuring absorbance at 280nm. Key concepts explained are molar extinction coefficient, absorbance spectra of biomolecules, and dilutions for preparation of solutions.
Spectrophotometry Lecture
This document discusses spectrophotometry techniques for measuring light absorption by molecules. It covers the electromagnetic spectrum, Beer-Lambert law, applications of UV-vis spectroscopy like determining cell density and protein concentration, and methods for measuring absorbance of molecules like DNA, RNA, proteins, and other biological compounds. Key concepts explained include the relationship between absorbance, molar extinction coefficient, concentration, and path length in the Beer-Lambert law.
Synthesis of 3-Substituted Coumarins by the Knoevenagel Condensation Reaction
The document summarizes a study on the synthesis of 3-substituted coumarins via the Knoevenagel condensation reaction of various 2-hydroxyaldehydes and malonate esters under solvent-free conditions using silica gel or basic alumina as catalyst. The reaction was carried out using microwave irradiation. Various coumarin products were obtained in moderate to good yields and characterized using techniques such as IR, NMR, and GC-MS. The method provides an improvement over traditional Knoevenagel condensation reactions through the use of solvent-free conditions and microwave activation.
dokumen.tips_-co-2-capture-using-deep-eutectic-solvents-prof-emadadeen-m-ali-...
This document discusses using deep eutectic solvents (DESs) for carbon dioxide (CO2) capture. It presents experimental results on CO2 solubility in various DESs composed of different salts and hydrogen bond donors. The CO2 solubility was found to depend on the salt type and salt-to-hydrogen bond donor ratio used in the DES. Models using the Peng-Robinson equation of state were able to predict CO2 solubility with reasonable accuracy when adjusting interaction parameters. Overall, DESs show potential as lower-cost and more sustainable alternatives to conventional solvents like monoethanolamine for CO2 separation applications.
eugenia-pechkova-university-of-genova-italy.ppt
The document discusses the use of Langmuir-Blodgett (LB) thin protein films for protein crystal growth using LB nanotemplates and robotics. Key points:
- LB technique is used to create organized thin films of proteins layer-by-layer at the air-water interface for use as nanotemplates in protein crystal growth.
- Various proteins including lysozyme, thaumatin, proteinase K, and ribonuclease A were tested using both classical and LB nanotemplate crystallization methods.
- LB nanotemplates were found to facilitate the nucleation and growth steps in crystallization, resulting in earlier formation and larger crystals compared to classical methods.
- Protein crystals
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Covalent Versus Electrostatic Attachment of Yeast Cytochrome c to a Fused Si...
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Surface Coverage Dependent Renaturation of Oxidized Cytochrome c Adsorbed to ...
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Modification & Application of Borate Zirconia Catalyst
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Jovanovic- VILNUS-E2a.pptx Kinetics of fularene polyhydroxylation
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Synthesis of 3-Substituted Coumarins by the Knoevenagel Condensation Reaction
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Study of the folding and unfolding of proteins adsorbed to a fused silica surface
- 1. Study of the Folding and Unfolding of Proteins Adsorbed to a Fused Silica Surface By Sheetal Mistry Department of Chemistry, Butler University Indianapolis, IN 46208
- 6. Cytochrome c Heme
- 8. General Function Voet, Donald; Voet, Judith; Pratt, Charlotte. Fundamentals of Biochemistry Upgrade Ed. 2001:501
- 16. Example Surface Spectra 1 µ M [YCC], 7mM Succinate Buffer, pH 4.00
- 17. Solution and Surface 1 µ M [YCC], 7mM Succinate Buffer, pH 4.00 Solution Surface
- 35. Surface vs. Solution: [Succinate Buffer] = 10 mM pH = 4.7 [NaCl] = 150 mM *Note the spectrum for 0.1 µM is enlarged Surface Adsorption
- 37. Surface vs. Solution: [Succinate Buffer] = 10 mM 60% n-propanol [NaCl] = 150 mM pH = 4.7
- 40. YCC Electrostatically attached
- 44. Is it really a monomer?...
- 45. Gel Electrophoresis HCC 1μg Dimer ~24,000g/mol Monomer~12,000g/mol YCC freshly dissolved 10 μ g YCC uncapped (6 mths) 1 μ g YCC uncapped (4 mths) 1 μ g YCC capped /purified 1 μ g
- 47. Encountered Problems at Step 2 Intensity proportional to Number of proteins on surface @ 409 nm
- 51. Determination of the [YCC] Abs max = 0.0054 pH 4.00, 7mM phosphate buffer Surface Adsorption of covalently anchored YCC
- 52. Determination of [YCC] ~0.0054 Covalently attached studies done at ~1.00 uM [YCC] Electrostatically adsorbed Surface Adsorption Isotherm
- 54. Alcohol Study on Surface Alcohol: 1 propanol pH 4.00 Buffer: 7 mM Succinate [YCC] : 1.00E-6M = 1.00 uM
- 55. Alcohol Study in Solution Alcohol: 1 propanol pH 4.00 Buffer: 7 mM Succinate [YCC] : 1.00E-6M
- 56. Surface and Solution Alcohol: 1 propanol pH 4.00 Buffer: 7 mM Succinate [YCC] : 1.00E-6M