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Structure of the Month

Structure of the Month: July 2009 [see all]

The consequences of disease-associated mutations on the structure and function of the filamin B actin binding domain

Gregory M Sawyer¹, Alice R Clark¹, Stephen P Robertson² and Andrew J Sutherland-Smith¹

Institute of Molecular BioSciences, Massey University, Private Bag 11222, Palmerston North 4410, New Zealand.
Department of Paediatrics and Child Health, Dunedin School of Medicine, University of Otago, PO Box 913, Dunedin 9054, New Zealand.

Filamin B (FLNB) crosslinks the F-actin cytoskeleton through an N-terminal actin binding domain (ABD) maintaining the structural integrity of cells and localises membrane receptors and signalling proteins. A cluster of missense mutations in the FLNB ABD is associated with human atelosteogenesis skeletal malformation disorders. To understand mechanisms of how these point mutations underlie disease we investigated the structure and function of two mutant domains (W148R and M202V) and compared their structures, stabilities and activities to the wild type domain.

All three structures display the classic compact monomeric conformation for the ABD showing that this conformation is stable to the sequence substitutions though changes in stability and actin binding activity are evident. Small local rearrangements are observed near the C-terminus for the W148R mutant but not M202V.

This work has been published in the Journal of Molecular Biology 2009 vol. 390 pp. 1030-1047.

Figure 1: Ribbon representation of the Filamin B actin binding domain colour ramped (blue to red) from N to C-terminus.

Figure 2: Superposition of the wild type (blue) and W148R (yellow) Filamin B actin binding domain structures. Wild type W148 and mutant R148 sidechains are shown in stick representation.

Figure 3: Superposition of the wild type (blue) and M202V (green) Filamin B actin binding domain structures. Wild type M202 and mutant V202 sidechains are shown in stick representation.

This research was performed in the laboratory of Andrew Sutherland-Smith in the Institute of Molecular BioSciences at Massey University Palmerston North, New Zealand.

Data collection details

Protein	FLNB WT	FLNB W148R	FLNB M202V
PDB ID	2WA5	2WA6	2WA7
Space group	P2₁2₁2₁	P2₁2₁2₁	P2₁2₁2₁
Unit cell	a = 43.05 Å b = 71.18 Å c = 90.44 Å	a = 46.03 Å b = 69.30 Å c = 89.27 Å	a = 46.08 Å b = 69.83 Å c = 87.18 Å
Radiation	Cu Kα	Cu Kα	Cu Kα
Generator	MicroMax-007	MicroMax-007	MicroMax-007
Optic	Blue	Blue	Blue
Detector	R-AXIS IV++	R-AXIS IV++	R-AXIS IV++
Crystal-to-detector distance	100 mm	140 mm	140 mm
Exposure time per frame	8 min	10 min	9 min
Oscillation width	0.25°	0.35°	0.15°
Number of frames	460	342	803
Data processing	Mosflm/Scala	Mosflm/Scala	Mosflm/Scala
Resolution range	20.3-1.90 Å	27.7-1.95 Å	27.9-1.85 Å

Protein 1: Human filamin B actin binding domain, PDB ID: 2WA5
Protein 2: W148R filamin B actin binding domain, PDB ID: 2WA6
Protein 3: M202V filamin B actin binding domain, PDB ID: 2WA7

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