The association state of a detergent-solubilized membrane protein measured during crystal nucleation and growth by small-angle neutron scattering

Reaction centers (RCs) from Rhodobacter sphaeroides R-26 were used as a model for membrane protein macromolecular organization and crystallization. Small-angle neutron-scattering measurements were made during pre-saturation, nucleation, and crystal growth phases of crystallization, using polyethylen...

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Bibliographic Details
Published in:Journal of crystal growth 1998-08, Vol.191 (4), p.811-819
Main Authors: Ann Marone, Palma, Thiyagarajan, P, Wagner, Arlene M, Tiede, David M
Format: Article
Language:English
Subjects:
Association reactions
Biological and medical sciences
Biological membranes
Crystal growth
Crystallization
Fundamental and applied biological sciences. Psychology
Macromolecules
Membrane protein
Molecular biophysics
Molecular structure
Neutron scattering
Nucleation
Polyethylene glycols
Reaction center
Saturation (materials composition)
Small-angle neutron scattering
Structure in molecular biology
Tridimensional structure
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Summary:Reaction centers (RCs) from Rhodobacter sphaeroides R-26 were used as a model for membrane protein macromolecular organization and crystallization. Small-angle neutron-scattering measurements were made during pre-saturation, nucleation, and crystal growth phases of crystallization, using polyethylene glycol, PEG, as the precipitant, and with fully deuterated reaction center protein. Solution 2H 2O/ 1H 2O ratios were adjusted to match the average scattering length density of the detergent, and allow the scattering from the deuterated reaction center to be selectively detected within the detergent and PEG containing mixtures. Neutron-scattering profiles for the detergent-solubilized reaction center in PEG -containing solutions were found not to deviate detectably from that expected for mono-dispersed, noninteracting particles. Analysis of the scattering in terms of discrete aggregates suggests that the reaction center exists predominately in the monomeric form throughout the crystallization process. The absence of detectable (
ISSN:0022-0248
1873-5002
DOI:10.1016/S0022-0248(98)00353-4