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Published by ogangurel

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Published by: ogangurel on Nov 18, 2008
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02/01/2013

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Crystallographic Studies on
Recombinant Human Stem Cell Factor \u00ad 141
17 November, 2008
Ogan Gurel
Prof. Wayne A. Hendrickson
Crystallization Experiments

The full\u00adlength extra\u00adcellular form of SCF is very soluble and after preliminary
screening trials, no crystals were obtained. Screening with the 141 amino acid\u00ad
long truncated form was more successful, however crystals that were ultimately
of the orthorhombic form were quickly obtained. When these crystals were first
observed, their complexity and uniqueness were not completely appreciated. As
it turns out, the conditions for crystallization are actually quite specific and
furthermore a number of crystal types may appear within the same droplet. The
sensitivity to conditions makes it difficult to systematically evaluate the
multiplicity of crystal types, while this multiplicity complicates the subsequent
diffraction analysis. Tables 1 & 2 summarize these results.

Photo # 1:
Orthorhombic crystals (long & block)
Photo # 2:
Hexagonal crystals
2
3
Table 1. rhSCF\u00ad141 Crystallization Conditions
Crystal Type
Initial crystallization
droplet
Equilibrium reservoir &
temperature
ORTHORHOMBIC
TypesA1,A2, &B
14.5 % PEG 400
12.5M CaCl2

50mM HEPES pH 7.5
50mM NaCl
5mM NaAcetate
5mg/ml rhSCF\u00ad141

29% PEG 400
0.25 M CaCl2
100mM HEPES pH 7.5
T = 22\u00b0C
HEXAGONAL
TypeC
0.29M Na.K phosphate
pH 9.0
0.29% PEG 400

57mM NaCl
5.7mM NaAcetate
5.7mg/ml rhSCF\u00ad141

2.0 M Na/K phosphate
T = 22\u00b0C

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