Rigid Body Refinement of Protein Complexes with Long-Range Distance Restraints from Pulsed Dipolar ESR.

Rigid Body Refinement of Protein Complexes with Long-Range Distance Restraints from Pulsed Dipolar ESR. Research Abstract Details 

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Rigid Body Refinement of Protein Complexes with Long-Range Distance Restraints from Pulsed Dipolar ESR. Abstract Text:

Jaya Bhatnagar,Jack H Freed,Brian R Crane,Jaya Bhatnagar,Jack H Freed,Brian R Crane,Jaya Bhatnagar,Jack H Freed,Brian R Crane,

The modeling of protein-protein complexes greatly benefits from the incorporation of experimental distance restraints. Pulsed dipolar electron spin resonance spectroscopy is one such powerful technique for obtaining long-range distance restraints in protein complexes. Measurements of the dipolar interaction between two spins placed specifically within a protein complex give information about the spin-spin separation distance. We have developed a convenient method to incorporate such long-range distance information in the modeling of protein-protein complexes that is based on rigid body refinement of the protein components with the software Crystallography and NMR System (CNS). Factors affecting convergence such as number of restraints, error allocation scheme, and number and position of spin labeling sites were investigated with real and simulated data. The use of 4 to 5 different labeling sites on each protein component was found to provide sufficient coverage for producing accuracies limited by the uncertainty in the spin-label conformation within the complex. With an asymmetric scheme of allocating this uncertainty, addition of simulated restraints revealed the importance of longer distances within a limited set of total restraints. We present two case studies: (1) refinement of the complex formed between the histidine kinase CheA and its coupling protein CheW, and (2) refinement of intra-helical separations in the protein a-synuclein bound to micelles.

Rigid Body Refinement of Protein Complexes with Long-Range Distance Restraints from Pulsed Dipolar ESR. Publishing Authors By Initials

J Bhatnagar,JH Freed,BR Crane,J Bhatnagar,JH Freed,BR Crane,J Bhatnagar,JH Freed,BR Crane,

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Rigid Body Refinement of Protein Complexes with Long-Range Distance Restraints from Pulsed Dipolar ESR. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Methods in enzymology

VOLUME: 423

Page Numbers: 117-33

Journal Abbreviation: Meth. Enzymol.

ISSN: 0076-6879

DAY: 4

MONTH: 07

YEAR: 2007

Rigid Body Refinement of Protein Complexes with Long-Range Distance Restraints from Pulsed Dipolar ESR. Information

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LANGUAGE: eng

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