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Nano-positioning system for structural analysis of functional homomeric proteins in multiple conformations.


ABSTRACT: Proteins may undergo multiple conformational changes required for their function. One strategy used to estimate target-site positions in unknown structural conformations involves single-pair resonance energy transfer (RET) distance measurements. However, interpretation of inter-residue distances is difficult when applied to three-dimensional structural rearrangements, especially in homomeric systems. We developed a positioning method using inverse trilateration/triangulation to map target sites within a homomeric protein in all defined states, with simultaneous functional recordings. The procedure accounts for probe diffusion to accurately determine the three-dimensional position and confidence region of lanthanide LRET donors attached to a target site (one per subunit), relative to a single fluorescent acceptor placed in a static site. As first application, the method is used to determine the position of a functional voltage-gated potassium channel's voltage sensor. Our results verify the crystal structure relaxed conformation and report on the resting and active conformations for which crystal structures are not available.

SUBMITTER: Hyde HC 

PROVIDER: S-EPMC3633233 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature

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Nano-positioning system for structural analysis of functional homomeric proteins in multiple conformations.

Hyde H Clark HC   Sandtner Walter W   Vargas Ernesto E   Dagcan Alper T AT   Robertson Janice L JL   Roux Benoit B   Correa Ana M AM   Bezanilla Francisco F  

Structure (London, England : 1993) 20121001 10


Proteins may undergo multiple conformational changes required for their function. One strategy used to estimate target-site positions in unknown structural conformations involves single-pair resonance energy transfer (RET) distance measurements. However, interpretation of inter-residue distances is difficult when applied to three-dimensional structural rearrangements, especially in homomeric systems. We developed a positioning method using inverse trilateration/triangulation to map target sites  ...[more]

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