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28 Cards in this Set
- Front
- Back
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What are the 5 properties of proteins that can be measured and exploited? |
Mass Charge-pH Hydrophobic/hydrophilic properties Differential solubility Mobility in applied fields |
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What is the largest protein in the body? What is its function and how many amino acid residues does it contain? |
Titin Present in muscle sarcomeres 33,000 amino acid residues |
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What is the smallest protein in the body? What is its function and how many amino acid residues does it contain? |
Sarcoplin Ca2+ transporter 31 amino acid residues |
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When discussing solvation shells, what do the following symbols correspond to: Rh, K^-1, Psi (r), Zeta? |
Rh: hydrodynamic radius K^-1: Debye length Psi (r): Electrostatic potential Zeta: Zeta potential |
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In liquid chromatography, there is a liquid moving phase. What two types of liquid chromatography are there? What do they have as a stationary phase? |
Liquid Solid chromatography: solid stationary phase Liquid Liquid chromatography: Liquid stationary phase |
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In gas chromatography, there is a gas moving phase. What two types of gas chromatography are there? What do they have as a stationary phase? |
Gas Solid chromatography: solid stationary phase Gas Liquid chromatography: Liquid stationary phase |
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Define a colloid. Define a sol. |
Colloid: Dispertion of small particles of one material in another Sol: Solid dispersal in a liquid |
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Define an aerosol. Define an emulsion. |
Aerosol: Liquid in a gas Emulsion: liquid in a liquid |
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Define a smoke. Define a lyophilic colloid. Define a lyophobic colloid. |
Smoke: Solid in a gas Lyophilic colloid: solvent attracting colloid Lyophobic colloid: Solvent repelling colloid |
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Define a gel. |
Semi-rigid mass of lyophilic sol in which the solvent has penetrated the sol particles |
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Gel filtration seperates by? |
Affinity of particles to gel particles |
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State 3 uses of gel filtration. |
-Fractionate proteins of different molecular weights -Remove aggregated protein from purified sample to produce homogenous protein sample -Desalt proteins |
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Sephadex is an example of a...? What is it used for? Sephadex has a large hydration shell, why is this? |
Cross-linked polysaccharide Separate low and high molecular weights Large number of Oxygen atoms |
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Sephacryl is a composite gel. What is it comprised of? Sepharose is a bead formed gel prepared from what? |
-Crosslinked allyl dextran with N,N'-methylene bisacrylamide -Agarose |
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Sephacryl and sepharose can operate under high pressures. What process are they used in? |
Fast Protein Liquid Chromatography |
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Parameters of gel filtration columns need to be controlled. Column length determines what 2 data properties? Do smaller or larger molecules pass through the gel the quickest? |
Resolution and elution time Larger |
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What is the relationship between Excluded Volume (Vo) and the volume of the column? Excluded Volume, along with MW, determines what? |
Vo is generally 1/3 of the coulmn volumn (so Vi is 2/3) The residence time of the protein |
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Ion exchange chromatography allows seperation based on what? |
Charge on protein molecule |
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Proteins bind mainly by electrostatic forces between the proteins surface charges and what? |
Dense clusters of charge groups on the ion exchange resin |
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Strong acidic cation exchange coulmns. What do they contain? What pH range do they work best in? They are useful in the fractionalisation of 5 compounds. State 3 of these compounds? |
Sulphonic acid group pH ranges 1-14 Answer can include any 3 of the following: cations, in organics, vitamins, peptides and amino acids |
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Weak acidic cation exchange columns. What do they contain? What pH range do they work best in? They are useful in the fractionalisation of 5 compounds. State 3 of these compounds. |
Carboxylic acid group pH range 5-14 Answer can include any 3 of the following: Biochemicals, cations, amino acids, antibiotics and organic bases |
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Strong basic anion exchange columns. What do they contain? What pH range do they work best in? They are useful in the fractionalisation of 3 compounds. State these 3. |
Quarternary ammonium groups pH range 0-12 Anions, vitamins and fatty acids |
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Weak basic anion exchange columns. They can contain 3 types of compound, state all 3. What pH range do they work best in? They are useful in the fractionalisation of 3 compounds. State these 3. |
Phenol, formaldehyde or polyamines pH 0-9 Anionic complexes of metals, vitamins and amino acids |
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Increasing the salt concentration elutes the stronger binding proteins from the columns. How is this acheived? |
Disruption of the solvation shell around the protein |
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In hydrophobic chromatography, the protein is usually bound at? What is used to provide this? How is the bound protein eluted from the column? |
High salt concentration Ammonium sulfate Reducing the salt gradient |
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Affinity chromatography separates proteins based on what? |
Basis of a reversable interaction between a protein and a specific ligand coupled to a chromotographic matrix |
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State 3 advantages of using affinity chromatography over other methods? |
-High selectivity -High capacity for protein -High purification of factor |
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5 types of componds can be used to seperate proteins in affinity chromotography. State 3 of these 5 compounds |
Answer can include any 3 of the following: Enzymes, glutathione, metal ions, antibodies or lectins |
