Growing Protein Crystals. Using Calcium-Integrin Binding Protein as a Model Presented by Chad Blamey. FBP www.scripps.edu/~arvai/ xtals/xtals.html. Goals. What are good crystals Why getting good crystals is important Understand how crystals grow
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Using Calcium-Integrin Binding Protein as a ModelPresented by
30% w/v Polyethelene glycol 5000
1 M NaCl
50 mM NaAcetate pH 4.5
50 mM Na Acetate pH 4.5
Should make large crystals in less than 15 minutes.
We will watch it for the hour of the lecture.
Poor, smeary spots
Notice ‘twined’ spots
Good! Round spots
Higher order visible (circle)
Do spots match mathematical predictions?
[Y]Growing Crystals: Hanging Drop Method
Growth & Nucleation
Barrier of Nucleation
Soluble proteinPhases of Proteins In Solution
Not to be confused with phases of light
CMCC figure 3.3
[Protein]Nucleation & Growth
[Y]Optimize Crystal Growth
For solution made up of three parts A, B and C.
Changing [C] will effect the quality of the crystal in
terms of [A].
0 hoursGrowing Crystals:Other Techniques
Specific Ions (Ca2+)
Relative Proportion of Conditions
Purity Of Protein*
Access to water*
Binding partnersFactors Effecting Crystal Growth
CIB purified w/o reducing agents
22CIB Protein Characterization
Further characterization of a protein can
improve purity and therefore crystal quality