Bioisis

"High-affinity interactions with the RING domain mediate stoichiometric GLMN binding to RBX1 complexes"

Experimental SAS Curve

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Low_res_thumbnail

Experimental Mass

91,000 Da

Experimental Details for BID:  GLOMNP
Experiment ID: 101
Collected at: ALS BL 12.3.1
Contributors: Hammel, M ,  Tainer, J ,  Weinkam, P ,  Schneidman-duhovny, D ,  Webb, B ,  Sali, A
Structural and biochemical analyses reveal that GLMN adopts a HEAT-like repeat fold that tightly binds the E2-interacting surface of RBX1, inhibiting CRL-mediated chain formation by the E2 CDC34. The structure explains the basis for GLMN’s selectivity toward RBX1 over RBX2, and how disease-associated mutations disrupt GLMN-RBX1 interactions. Our study reveals a mechanism for RING E3 ligase regulation whereby an inhibitor blocks E2 access, and raises the possibility that other E3s are likewise controlled by cellular proteins that mask E2-binding surfaces to mediate inhibition.
single concentration

Electron Pair Distribution

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       Dmax → 127 Å


Guinier Plot

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     Guinier Rg → 38.15 Å

Real Space Rg → 40.95 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.


Ensemble Model

A ENSEMBLE model was determined using the following:

Simulation MethodAllosMod-FoXS 
Simulation AlgorithmMD 
Ensemble Size10000 
Selection MethodMES 
Member Size
Scoring Functionchi 
Score0.1 

Ensemble Fit

Med_res_ensemble_fit

The red line is the calculated SAXS profile from the ENSEMBLE model scaled to the experimental SAXS curve (cyan).


Additional Experimental Details
Title

High-affinity interactions with the RING domain mediate stoichiometric GLMN binding to RBX1 complexes

Description

Structural and biochemical analyses reveal that GLMN adopts a HEAT-like repeat fold that tightly binds the E2-interacting surface of RBX1, inhibiting CRL-mediated chain formation by the E2 CDC34. The structure explains the basis for GLMN’s selectivity toward RBX1 over RBX2, and how disease-associated mutations disrupt GLMN-RBX1 interactions. Our study reveals a mechanism for RING E3 ligase regulation whereby an inhibitor blocks E2 access, and raises the possibility that other E3s are likewise controlled by cellular proteins that mask E2-binding surfaces to mediate inhibition.

Publication

Structure of a glomulin-RBX1-CUL1 complex: inhibition of a RING E3 ligase through masking of its E2-binding surface, Molecular Cell, Vol. 47(3), 371-382

Contributors

Hammel, M ,  Tainer, J ,  Weinkam, P ,  Schneidman-duhovny, D ,  Webb, B ,  Sali, A

Genomics and Proteomics

The experiment is composed of a single gene/ORF

Abbreviated name: GLOMULIN

Annotation: Monomer. Isoform 1 interacts with non-phosphorylated MET and is released upon receptor phosphorylation. Isoform 2 interacts with FKBP59 and FKBP12. Isoform 1 is part of a SCF-like complex consisting of CUL7, RBX1, SKP1, FBXW8 and GLMN isoform 1.

MAVEELQSII KRCQILEEQD FKEEDFGLFQ LAGQRCIEEG HTDQLLEIIQ NEKNIVIIKN MGWNLVGPVV RCLLCKDKED SKRKVYFLIF DLLVKLCNPK ELLLGLLELI EEPSGKQISQ SILLLLQPLQ TVIQKLHNKA YSIGLALSTL WNQLSLLPVP YSKEQIQMDD YGLCQCCKAL IEFTKPFVEE VIDNKENSLE NEKLKDELLK FCFKSLKCPL LTAQFFEQSE EGGNDPFRYF ASEIIGFLSA IGHPFPKMIF NHGRKKRTWN YLEFEEEENK QLADSMASLA YLVFVQGIHI DQLPMVLSPL YLLQFNMGHI EVFLQRTEES VISKGLELLE NSLLRIEDNS LLYQYLEIKS FLTVPQGLVK VMTLCPIETL RKKSLAMLQL YINKLDSQGK YTLFRCLLNT SNHSGVEAFI IQNIKNQIDM SLKRTRNNKW FTGPQLISLL DLVLFLPEGA ETDLLQNSDR IMASLNLLRY LVIKDNENDN QTGLWTELGN IENNFLKPLH IGLNMSKAHY EAEIKNSQEA QKSKDLCSIT VSGEEIPNMP PEMQLKVLHS ALFTFDLIES VLARVEELIE IKTKSTSEEN IGIK
categoryamino acid composition(%)
HydrophobicI(7.9) V(4.4) L(16.0) M(2.4) A(3.4) G(4.5) P(3.5)
AromaticF(4.7) W(0.8) Y(2.4)
HydrophilicR(2.7) K(7.7) E(9.8) D(3.9) Q(5.9) N(6.1) H(1.7) S(6.6) T(3.5) C(2.2)