Structural Differences

Title: Separation Lab Purpose: The purpose of this lab was to successfully separate a mixture of iron filings, poppy seeds, sand and salt. Hypothesis: In my opinion, I think it is possible to successfully separate the mixture.

This Lab Report is an analysis of the results of a two-part experiment. In the first part, we used a gel filtration column to separate the components of a mixture composed of protein and non-protein molecules. By doing so we hoped to obtain fractions that contained single components of the mixture, while also gaining insight into the relative molecular weight of each component compared to each other. We would then plot these fractions onto nitrocellulose paper in order to determine which fractions had protein. In the second part, we would use the fractions which we had determined had protein to conduct an SDS-PAGE. By doing so we hoped to determine an estimate on the molecular weight of the proteins present in each fraction by comparing it to a tracker dye composed of a variety of molecules of differing molecular weight.

When we hear the chromatography we think of many different things. Chromatography is referred to as several related techniques to separate mixtures of compounds, analyze and identify. Although there are different techniques the one thing that they all have in common is that they have an operation that is two parts. The two parts are known as mobile phase and stationary phase. In the mobile phase is known as the liquid or gas that the mixture is placed in.

The objective of this extraction experiment was to achieve a comprehensive understanding, as well as master the practice, of the technique of separating various individual components of a compound.

The purpose of this lab is to separate a mixture and determine the percentages of each of the ingredients. Each substance will have a different boiling point due to its intrinsic properties and from that, we will be able to determine the purity of different products as we evaporate off the next level of product.

The stationary phase will absorb or slow down different components of the tested solution to different degrees creating layers as the components of the solution are separated. Chromatography was invented by the Russian botanist, Mikhail Tsvet. Chemists use this process to identify unknown substances by separating them into the different molecules that make them up.

-terminal fragment of the YFP was fused to the N-terminus of TuHC and AtCA1 to make YFPN-TuHC and YFPN-AtCA1, respectively, while the C-terminal fragment of the YFP was fused to the N-terminus of TuHC and TuMV CP (TuCP)

Proteins are polymeric chains that are built from monomers called amino acids. All structural and functional properties of proteins derive from the chemical properties of the polypeptide chain. There are four levels of protein structural organization: primary, secondary, tertiary, and quaternary. Primary structure is defined as the linear sequence of amino acids in a polypeptide chain. The secondary structure refers to certain regular geometric figures of the chain. Tertiary structure results from long-range contacts within the chain. The quaternary structure is the organization of protein subunits, or two or more independent polypeptide chains.

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In many cases, the proteins in question are monoclonal antibodies generated against a particular target of interest such as a cell surface receptor, a cytoskeletal protein, or a cellular organelle. In some cases, a fluorescent label is used to track the fate of a particular protein as it is processed in the ER and Golgi complex, transported along microtubules, or secreted from the cell altogether. In still other cases, two or more fluorescently tagged proteins are introduced into the same cell to see whether they localize in the same or separate compartments or organelles.

In their article, “Differences in Mouse Maternal Care Behavior – Is There a Genetic Impact of the Glucocorticoid Receptor,” Chourbaji et al. (2011) report a 2*2 factorial research design to determine the effect of mutations in Glucocorticoid Receptor (GR) on the maternal behaviors in two different strains of inbred mice. The experiment was conducted in three main stages. The first stage was the production of GR heterozygous mice of both lab strains under study. The next step was the controlled mating and delivery of the pups for knowing the effect of this mutation. The third step was the observation of the maternal behavior in two strains. No significant differences in the behaviors of two strains were observed and

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

G-proteins are have three subunits, alpha (), beta () and gamma (), each composed of different amino acid therefore they are known as heterotrimeric. The G and G are tightly attached to each other so are known as the beta-gamma complex (G). G subunit has an important feature which is the binding site for exchange of GTP to GDP.

Campbell and Farrell define proteins as polymers of amino acids that have been covalently joined through peptide bonds to form amino acid chains (61). A short amino acid chain comprising of thirty amino acids forms a peptide, and a longer chain of amino acids forms a polypeptide or a protein. Each of the amino acids making up a protein, has a fundamental design that comprises of a central carbon or alpha carbon that is bonded to a hydrogen element, an amino grouping, a carboxyl grouping, and a unique side chain or the R-group (Campbell and Farrell 61).

SDS-PAGE and Western blot 39 4. Results 41 4.1. How does UPF1 bind its substrates? 41 4.1.1. Introduction 41 4.1.2.