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Calculation of generalized structural parameters (Dmax, Rg) Determination of the molecular shape of macro-molecules p Differentiation of mono-disperse and aggregated solutions Differentiation of folded and unfolded protein solutions Characterization of oligomeric states
Log(I)
Integrated profile
q [-1]
Complimentary techniques
Crystallography Single crystals High resolution (up to 0.1nm) Atomic t t At i structure information i f ti Crystal required
SAXS Dilute solutions (1 ~ 100mg/ml) Analysis in native conditions Low resolution (~1-2nm) Modeling ambiguity
Scattering intensity
I ( q) A( q) A * ( q)
X-ray diffraction
Ahkl (q ) = f j exp{2i ( hx j + ky j + lz j )}
j =1 N
z y x r
Patterson function
P(r)
r
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Profile conversion
Log(I) )
SAXS pattern
q [-1]
Guinier plot
ln n(I) q2I
Kratky plot
P P(r)
q2 [-2]
q [-1]
r []
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Guinier plot
3 q 2 RG ln[ I ( q)] = ln[ I (0)] 3
Mono-disperse
Aggregated
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Kratky plot
Folded
Partially unfolded
Unfolded
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RG 2 =
r P P ( r ) dr ( r ) dr
2 0 0
Dmax
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~3m
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MicroMax 007
MM007
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MicroMax 002+
MM002+
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Sample feeder
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Rg = 132 04 132.04
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Kratky plot
HSA (human serum albumin) Well folded
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Rg = 29 02 29.02
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Review papers
Robust, high throughput Robust high-throughput solution structural analyses by small angle X-ray scattering (SAXS) Greg L Hura et al., Nature Methods, July 20 (2009) Small-angle scattering studies of biological macromolecules in solution Dmitri I Svergun et al Rep Prog Phys 66 1735 (2003) al., Rep. Prog. Phys. X-ray solution scattering (SAXS) combined with crystallography and computation Christopher D Putnam et al., Q. Rev. Biophys. 40, 191 (2007)
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Reference book
S ll Small-angle scattering of X-rays l tt i fX Andre Guinier & Gerard Fournet Wiley Wil (1955) ( t f i t) (out-of-print) ProQuest www.umi.com
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Conference
Th 67th Annual Pittsburgh Diffraction Conference The A l Pitt b h Diff ti C f October 29th 31st University of Georgia Center for Continuing Education http://www.pittdifsoc.org/PDC_2009/
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Thank you ! a
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