Bioisis

"Glucose Isomerase under different ionic strengths using NaCl concentrations varying between 0 and 1000mM"

Experimental SAS Curve

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Experimental Mass

174,000 Da

Experimental Details for BID:  GNaClP
Experiment ID: 60
Collected at: ALS BL12.3.1
Contributors:
SAXS profiles for glucose isomerase and lysozyme at the concentration of ~2.5 mg/ml were collected under different ionic strengths using NaCl or KCl concentrations varying between 0 and 1000mM. The SAXS data for contrast variation experiments have been collected using two standard protein glucose isomerase (174 kDa). Protein at the concentration 5mg/ml were first dialyzed in buffer containing 50mM Tris pH 7.6. Dialyzed proteins were aliquot into the 50uL fractions that were diluted with 50uL of different NaCl salt solutions giving final salt concentration varying between 25mM and 1000mM. The corresponding buffer blanks for SAXS experiments were prepared identically by diluting 50uL aliquots of the dialyzing buffer with salt solutions. SAXS data was collected and analyzed as described previously (Hura, 2009 Nature Method).
SAXS profiles for glucose isomerase and lysozyme at the concentration of ~2.5 mg/ml were collected under different ionic strengths using NaCl or KCl concentrations varying between 0 and 1000mM. The SAXS data for contrast variation experiments have been co

Electron Pair Distribution

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       Dmax → 97 Å


Guinier Plot

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     Guinier Rg → 31.0 Å

Real Space Rg → 31.0 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 4.0.


Additional Experimental Details
Title

Glucose Isomerase under different ionic strengths using NaCl concentrations varying between 0 and 1000mM

Description

SAXS profiles for glucose isomerase and lysozyme at the concentration of ~2.5 mg/ml were collected under different ionic strengths using NaCl or KCl concentrations varying between 0 and 1000mM. The SAXS data for contrast variation experiments have been collected using two standard protein glucose isomerase (174 kDa). Protein at the concentration 5mg/ml were first dialyzed in buffer containing 50mM Tris pH 7.6. Dialyzed proteins were aliquot into the 50uL fractions that were diluted with 50uL of different NaCl salt solutions giving final salt concentration varying between 25mM and 1000mM. The corresponding buffer blanks for SAXS experiments were prepared identically by diluting 50uL aliquots of the dialyzing buffer with salt solutions. SAXS data was collected and analyzed as described previously (Hura, 2009 Nature Method).

Publication

Contributors

Genomics and Proteomics

The experiment is composed of 2 genes/ORFs

Abbreviated name: GISRUB

Annotation: Xylose. isomerase. from. Streptomyces. rubiginosus

NYQPTPEDRF TFGLWTVGWQ GRDPFGDATR RALDPVESVR RLAELGAHGV TFHDDDLIPF GSSDSEREEH VKRFRQALDD TGMKVPMATT NLFTHPVFKD GGFTANDRDV RRYALRKTIR NIDLAVELGA ETYVAWGGRE GAESGGAKDV RDALDRMKEA FDLLGEYVTS QGYDIRFAIE PKPNEPRGDI LLPTVGHALA FIERLERPEL YGVNPEVGHE QMAGLNFPHG IAQALWAGKL FHIDLNGQNG IKYDQDLRFG AGDLRAAFWL VDLLESAGYS GPRHFDFKPP RTEDFDGVWA SAAGCMRNYL ILKERAAAFR ADPEVQEALR ASRLDELARP TAADGLQALL DDRSAFEEFD VDAAAARGMA FERLDQLAMD HLLGAR
categoryamino acid composition(%)
HydrophobicI(2.8) V(4.9) L(10.1) M(1.8) A(12.2) G(9.3) P(4.9)
AromaticF(6.0) W(1.6) Y(2.3)
HydrophilicR(9.1) K(2.8) E(7.3) D(9.6) Q(2.8) N(2.6) H(2.6) S(2.8) T(4.1) C(0.3)

Abbreviated name: MISTAKE

Annotation: Not in Experiment

NOT IN EXP ERIMENT
categoryamino acid composition(%)
HydrophobicI(11.8) V(0.0) L(0.0) M(5.9) A(0.0) G(0.0) P(5.9)
AromaticF(0.0) W(0.0) Y(0.0)
HydrophilicR(5.9) K(0.0) E(17.6) D(0.0) Q(0.0) N(17.6) H(0.0) S(0.0) T(11.8) C(0.0)