Bioisis

"Fusion Protein of Folded PF1282 and unfolded PF1205"

Experimental SAS Curve

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Experimental Mass

24,262 Da

Experimental Details for BID:  RBFSHP
Experiment ID: 3
Collected at: ALS BL 12.3.1
Contributors: Rambo, R.P. ,  Hura, G.L. ,  Menon, A.L. ,  Hammel, M. ,  Adams, M.W.W. ,  Tainer, J.A.
Part of a set of samples prepared recombinantly by Mike Adams group in Georgia. Has his-tag attached see sequence. This is a fusion protein of red rubredoxin to PF1205 which did not express alone in E. coli. The rubredoxin improved solubility and was a marker for expression. However, analysis suggests PF1205 is unfolded. Data is not on an absolute scale and is a compilation of several data sets collected at different wavelengths.

Electron Pair Distribution

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       Dmax → 95 Å


Guinier Plot

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     Guinier Rg → 24.0 Å

Real Space Rg → 25.0 Å

The Guinier plot is used to estimate the radius of gyration, Rg, which is taken from the slope of a line observed at low scattering angles (typically in the range where q* Rg < 1.3). This should be in reasonable agreement with the real space Rg.


Kratky Plot

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The Kratky plot can be used to visually assess the degree of "unfoldedness" of a protein or RNA sample. The plot of a well-behaved folded protein approaches the baseline at high q values creating a parabolic shape.


PDB Model fit to SAXS Data

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The red line is the calculated SAXS profile from a PDB model scaled to the experimental SAXS curve (cyan). The two curves agree with a Chi2 of 3.1.


DAMMIN/F Model

DAMMIN/F result was determined with the following:

Space GroupP1
NSD | RMSD0.629 
variance(NSD | RMSD)0.1 
Number of Models in Average10 

GASBOR Model

GASBOR result was determined with the following:

Space GroupP1
Chi5.76
NSD | RMSD0.89 
variance(NSD | RMSD)0.1 
Number of Models in Average10 
Superimposed model10 

Additional Experimental Details
Title

Fusion Protein of Folded PF1282 and unfolded PF1205

Description

Part of a set of samples prepared recombinantly by Mike Adams group in Georgia. Has his-tag attached see sequence. This is a fusion protein of red rubredoxin to PF1205 which did not express alone in E. coli. The rubredoxin improved solubility and was a marker for expression. However, analysis suggests PF1205 is unfolded. Data is not on an absolute scale and is a compilation of several data sets collected at different wavelengths.

Publication

Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS). Nat Methods. 2009 Aug;6(8):606-12.

Contributors

Rambo, R.P. ,  Hura, G.L. ,  Menon, A.L. ,  Hammel, M. ,  Adams, M.W.W. ,  Tainer, J.A.

Genomics and Proteomics

The experiment is composed of 2 genes/ORFs

Abbreviated name: PF1205

Annotation: hypothetical protein

MKLVLDSSVF IQGVEVEGYT TPSVVEEIKD RESKLFLESL ISAGKVRVIE PSKEAIEKII QAARETGELE ELSKADIEVL ALAYELKATI FTDDYNVQNI ASLLGLKFRT LKKGITRVMK WRYVCIGCGR KFSTLPPGGI CPDCGSRVKL IPRRR
categoryamino acid composition(%)
HydrophobicI(9.0) V(8.4) L(10.3) M(1.3) A(5.8) G(7.1) P(3.9)
AromaticF(3.2) W(0.6) Y(2.6)
HydrophilicR(7.1) K(9.0) E(9.7) D(3.9) Q(1.9) N(1.3) H(0.0) S(7.1) T(5.2) C(2.6)

Abbreviated name: PF1282

Annotation: rubredoxin

MAKWVCKICG YIYDEDAGDP DNGISPGTKF EELPDDWVCP ICGAPKSEFE KLED
categoryamino acid composition(%)
HydrophobicI(7.4) V(3.7) L(3.7) M(1.9) A(5.6) G(9.3) P(9.3)
AromaticF(3.7) W(3.7) Y(3.7)
HydrophilicR(0.0) K(9.3) E(11.1) D(13.0) Q(0.0) N(1.9) H(0.0) S(3.7) T(1.9) C(7.4)