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21 Cards in this Set
- Front
- Back
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All proteins show physiological interactions with other molecules. State 4 examples of what there other molecules could be? |
Answer can include any 4 of the following: Other proteins Other macromolecules Signals Substrates Inhibitors |
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State 2 reasons why it is important to study protein interactions. |
1. Many diseases are the result of aberrant protein-protein interactions 2. Show how complexes work-drive biotechnology |
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All interactions have 4 key characteristics. State all 4 of these. |
Qualitatively Dissociation constant Stoichiometry On/Off rates |
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State the equation used to define the dissociation constant, Kd? |
Kd=[A]x[B]y/[AxBy] |
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The methods that we use to determine protein-protein interactions depends on 5 key issues. State 4 of these issues. |
Answer can include any 4 of the following: Purity of protein Amount of sample Prior knowledge Value of information Nature of the protein |
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There are more than 10 methods we can use to determine the protein-protein interactions. State 7 of these. |
Answer can include any 7 of the following: ELISA, Immuno/radio absorption, Fluorescence, Surface Plasmon Resonance, Isothermal Titration Calorimetry, Ultracentrifugation, Immunoprecipitation, Differential Scanning Fluorimetry, Co-Purification, Non-Dissociation Mass Spectronomy and Bilayer Interferometry |
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Why isn't the stoichiometric values always 1:1? |
Several ligands can bind to 1 receptor |
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What do the On/Off Rates refer to? |
Ability of the proteins to associate and dissociate |
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What does MIP stand for, it is given as an example. What is its function? |
Macrophage Infectivity Potentiator Chaperone for Burkholderia pseudomallei |
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State 2 disadvantages of preparing BpMip. |
Compounds are experimental and expensive |
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What is the use of Cetuximab? What does it target? |
Therapeutic antibody used to treat cancers Targets epidermal growth factor receptor 1 |
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State 1 positive and 1 negative of investigating protein-protein interactions using Cetuximab. |
EGFR preparation is easy Variant antibody production is difficult |
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What is BRCA2 involved in? What does it coordinate? |
DNA repair by homologous recombination Recombinase Rad51 to start homologous recombination through its 8 'BRCA2 repeat' regions |
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State a positive and a disadvantage of investigating protein-protein interactions using Cetuximab. |
Rad51 is easy to prepare BRCA2 is difficult to prepare |
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In fluorescence, the absorbed light can be pre-polarised before being shone on the sample. The time delay between absorbtion and emission, emission may happen in another orientation. This process is called? What 2 types of molecules can it distinguish between? |
Fluorescence anisotropy Fast tumbling and slow tumbling |
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The intensity of fluorescent light is measured in what 2 orientations? |
Parallel and perpendicular polarisations |
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What determines tumbling rate? What, in turn, determines this^? What can be done to reduce the tumbling rate? What affect does this^ have on the fluorescence anisotropy value? |
Brownian Motion Size of the protein Bind the protein to a larger molecule Reduces it |
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State the equation used to calculate Fluorescence Anisotropy. |
R= I=-III/I=+2III |
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A common approach in FA is to use _ of a labelled marker as a signal. |
Displacement |
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State 4 advantages of using FA as a technique to investigate protein-protein interactions. |
Answer can include any4 of the following: Versatile technique applicable to many problems Uses an instrument that can be used in other experiments High throughput Flexibility in solvents Moderate quantities required |
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State 4 disadvantages of using FA as a technique to investigate protein-protein interactions. |
Must have a fluorescent label Only gives KD Instrument not always available Usually need to get the added compound to 5x KD, may be difficult |
