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Quiz

Lecture 3

Kate H 12 Fragen

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  1. 1Why xray crystallography?
    - to determine [blank_start]spatial[blank_end] arrangement of atoms (size, bond length, bond angle, bond type)
    Define mechanistic details of binding and [blank_start]catalysis[blank_end]
    Elucidate structural and [blank_start]functional[blank_end] relationship - have structure and use biochemical knowledge. [blank_start]NMR[blank_end] common technique to obtain structural info.
    Structure-based drug design - method of designing medicines that relies on [blank_start]computational[blank_end] models of protein structures.
    1. ANMR
    2. Bcomputational
    3. Cspatial
    4. Dfunctional
    5. Ecatalysis
  2. 2How to induce crystal formation.

    [blank_start]Supersaturation[blank_end] - state of solution that contains more of the [blank_start]dissolved[blank_end] material than could be dissolved by the [blank_start]solvent[blank_end] under normal conditions.
    Water evaporation
    [blank_start]Nucleation[blank_end] - first step in formation. First contact that occurs between atoms that leads to further atom contact.
    [blank_start]Crystal[blank_end] formation - drop in temp causes falling of solution in an ordered way to form a crystal.
    1. ASupersaturation
    2. BNucleation
    3. CCrystal
    4. Ddissolved
    5. Esolvent
  3. 3Why is crystallisation of macromolecules difficult?

    - complex, highly mobile surfaces, heterogeneity
    - [blank_start]electrostatic[blank_end] nature - uneven [blank_start]charge[blank_end] distribution
    - [blank_start]low[blank_end] chemical and [blank_start]physical[blank_end] stability - unfolding, hydration, temp <[blank_start]40[blank_end] proteins begin to lose structure
    - overproduction, concentrating, purity
    - specific factors - [blank_start]prostethic[blank_end] groups and ligands
    1. Acharge
    2. B40
    3. Cphysical
    4. Dprostethic
    5. Eelectrostatic
    6. Flow
  4. 4Gene structure pipeline

    Genomic info > Identification of [blank_start]genes[blank_end] > [blank_start]Cloning[blank_end] > Expression? [blank_start]Activity[blank_end]
    1. Agenes
    2. BCloning
    3. CActivity
  5. 5Structural Genomics

    Depend on [blank_start]expression[blank_end] [blank_start]vector[blank_end] choice
    Aim to unravel structural functional relationship of all proteins encoded by [blank_start]genomes[blank_end]
    They're high [blank_start]throughput[blank_end]
    Limited by protein purification [blank_start]yields[blank_end]
    [blank_start]Purification[blank_end] tag-position affects outcomes of process
    1. Aexpression
    2. Bvector
    3. Cthroughput
    4. Dyields
    5. EPurification
    6. Fgenomes
  6. 6Important factors in crystallisation

    - type of salt and/or salt [blank_start]conc[blank_end]
    - type of salt and/or type of [blank_start]buffer[blank_end] pH
    - [blank_start]colloidal[blank_end] properties of proteins, ionic strength and pH
    - metal ions - solve [blank_start]phasing[blank_end] problems
    - [blank_start]precipitant[blank_end] - some in orderly fashion, some completely [blank_start]amorphouss[blank_end]
    1. Aconc
    2. Bcolloidal
    3. Cphasing
    4. Damorphouss
    5. Eprecipitant
    6. Fbuffer
  7. 7A crystal is a group of [blank_start]molecules[blank_end] arranged in a [blank_start]neat[blank_end] and orderly fashion. In an ideal crystal, content of each unit cell is [blank_start]identical[blank_end]. Precise stacking of unit cells allows crystal to have sharp [blank_start]edges[blank_end] and flat [blank_start]faces[blank_end].

    A crystal can be thought of being made u p of many different sets of [blank_start]equidistant[blank_end] parallel planes passing through the lattice points - lattice planes. Many properties of the crystal can be understood in terms of spatial geometry of these planes. [blank_start]Miller[blank_end] [blank_start]indices[blank_end] are used to designate these different sets of planes.

    How do we choose unit cell?
    - [blank_start]simplest[blank_end] (smallest) repeating unit
    - has to reflect [blank_start]symmetry[blank_end] of the crystal
    -symmetrical operations are translation, rotation, reflection, inversion, glide
    1. Amolecules
    2. Bneat
    3. Cidentical
    4. Dedges
    5. Efaces
    6. Fequidistant
    7. GMiller
    8. Hindices
    9. Isimplest
    10. Jsymmetry
  8. 8Precipitation zone - [blank_start]aggregation[blank_end]
    [blank_start]Nucleation[blank_end] zone - crystalline form - encourages [blank_start]crystal[blank_end] formation
    [blank_start]Metastable[blank_end] zone - [blank_start]supersaturation[blank_end] - can physically affect crystal formation
    1. ANucleation
    2. BMetastable
    3. Csupersaturation
    4. Dcrystal
    5. Eaggregation
  9. 9Transitions in crystal formation:
    Precipitate
    Clear drop
    Phase separatin
    [blank_start]Granular[blank_end] precipitate
    Spherulites
    Cluster of [blank_start]needles[blank_end]
    Plates
    [blank_start]Microcrystals[blank_end]
    Non-defined edges
    Crystals from [blank_start]heavy[blank_end] precipitate
    Crystals from granular precipitate
    [blank_start]Optimised[blank_end] crystals
    1. AGranular
    2. BMicrocrystals
    3. Cneedles
    4. Dheavy
    5. EOptimised
  10. 10Vapor diffusion set up:
    [blank_start]Difference[blank_end] in precipitant concentration drives water evaporation. Protein goes from unsaturation to [blank_start]supersaturation[blank_end].
    Hangin drop and sitting drop.
    Precipitant solution contains precipitant ([blank_start]ammonium[blank_end] sulphate), salt (NaCl), [blank_start]buffer[blank_end] (Tris-HCl pH 7.5)

    Volume of reservoir solution [blank_start]same[blank_end] as volume of protein
    Concentration of reservoir is [blank_start]double[blank_end] conc of reservoir in drop so drives [blank_start]evaporation[blank_end], keeps evaporating until same conc, protein conc increases unsat to supersat.

    If left for long, quality of crystal deteriorates.
    1. Asame
    2. Bdouble
    3. CDifference
    4. Dbuffer
    5. Eammonium
    6. Fevaporation
    7. Gsupersaturation

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Beschreibung
Biochemistry Quiz am Lecture 3, erstellt von Kate H am 01/04/2018.
Autor
Kate H
Sprache
English
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0
Schlagwörter
biochemistry
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