Protein Chromatography Essay

Proteins are an essential component of the human body tissues. They are made by combining amino acids together. There are many pros as well as cons to consuming inadequate amounts of protein each day. Making sure to consume the accurate type and amount of protein is important for the body. There are specific groups that do require more protein, however, if you are not a part of one of these groups consuming too much protein can have harmful effects.

The most common macromolecules found in living organisms are lipids, carbohydrates, proteins and nucleic acids. Briefly, the meaning of macromolecules is that they normally contain two or more molecules in them and their main functions are to store energy, information and much more. Most foods are known to be combinations of macromolecules. While some of these compounds can be detected by taste tests, many cannot. Scientists then use certain tests to determine the presence of macromolecules.

Adsorption chromatography occurs when “one substance form[s] some sort of bonds to the surface of another one”, creating intermolecular forces between the two substances (Chemguide). For thin layer chromatography, the components of the mixture are adsorbed onto the stationary phase that covers the plate (Chemguide). The more polar a substance is, the more strongly adsorbed it is (Chemguide) and the strong intermolecular forces then result in a slower rate of migration with respect to the moving phase. As the solvent touches the TLC plate, the solute (mixture) is allowed to move up the TLC plate

Which one of the statements concerning valence bond (VB) and molecular orbital (MO) bond theories is correct?

In this investigation concentration of protein was to be found using the concentration of a known protein( already labeled) by measuring the concentration of protein in milk and the absorbance of light. It is hypothesized that the protein that will be tested will be the same as the already labeled one. The outcome from the lab shows that the protein that was used did not respond the same as the already labeled one known from the standard curve, therefore rejecting the hypothesis.The results prove the hypothesis as incorrect because the proteins concentration were different when the concentration was calculated and all had a high percent difference.

In this column chromatography, acetylferrocene was more polar, therefore was held by the silica gel more tightly and moved through the column more slowly when the eluting solvent was nonpolar hexane. Increasing the polarity of the solvent would move all components faster. That explained when the solvent was switched to the more

“Enzymes are proteins that have catalytic functions” [1], “that speed up or slow down reactions”[2], “indispensable to maintenance and activity of life”[1]. They are each very specific, and will only work when a particular substrate fits in their active site. An active site is “a region on the surface of an enzyme where the substrate binds, and where the reaction occurs”[2].

Protein Synthesis Protein Synthesis is the process whereby DNA (deoxyribonucleic acid) codes for the production of essential proteins, such as enzymes and hormones. Proteins are long chains of molecules called amino acids. Different proteins are made by using different sequences and varying numbers of amino acids. The smallest protein consists of fifty amino acids and the largest is about three thousand amino acids long. Protein synthesis occurs on ribosomes in the cytoplasm of a cell but is controlled by DNA located in the nucleus.

2. How do the products of alternative splicing differ and how are they similar? (1 pt)

In recent years, interest has sparked the health industry to research protein consumption after exercise to optimise muscle growth. Muscle Protein Synthesis (MPS) is not only an appealing topic for bodybuilders, it is also beneficial to those over 60 years of age who may have experienced muscle mass loss over time. A decrease in muscle mass does not only lead to a decrease in overall strength but also makes daily movement severely painful, leading to a poor quality of life. All individuals can benefit from increasing muscle protein synthesis via an increased protein intake after resistance training. This will aid in maintaining muscle mass, strength and overall health (Wolfe, 2012).

     In liquid chromatography, the separator is called the column and consists in most cases of a tube filled with porous material called the stationary phase. A liquid, called the mobile phase, flows through the tube between the particles of stationary phase material. A liquid sample is taken from a mixture to be analyzed and introduced to a part of the system that is at elevated pressure. The sample is then transported to a separator by the flow in the system.

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

PEGylation is the chemical reaction wherein poly (ethylene glycol) or (PEG) is covalently conjugated to a protein. The resulting protein will attain improved pharmacokinetics when compared to the native protein such as increased in vivo half-life, decrease immunogenicity, increased hydrophilicity etc. The objective of this experiment is to PEGylate the protein lysozyme (lys) and study the kinetics of the PEG-lys when treated with the protein digestive enzyme trypsin. Designing generic therapeutic is currently on the forefront of Pharmaceutical biotechnology. One attractive technique in this area is PEGylation.

Chromatography Investigation Chromatography is a highly regarded technique used to separate the components of a mixture. It is based on the principle that each component possesses a unique affinity for a stationary phase and a mobile phase. The components that are more inclined to enter the mobile phase will migrate further on the chromatogram and distinguish themselves from the other components. The type of solvent used in chromatography is known to directly affect the separation of the mixture. In this experiment, thin-layer and column chromatography will be utilized to separate the numerous chlorophyll and carotenoid pigments of a spinach extract.

The A0 urine sample comes from a 30-year-old adult who is also an extreme athlete.