title: The membrane protein LeuT in micellar systems: aggregation dynamics and detergent binding to the S2 site.
abstract: Structural and functional properties of integral membrane proteins are often studied in detergent micellar environments (proteomicelles), but how such proteomicelles form and organize is not well understood. This makes it difficult to evaluate the relationship between the properties of the proteins measured in such a detergent-solubilized form and under native conditions. To obtain mechanistic information about this relationship for the leucine transporter (LeuT), a prokaryotic homologue of the mammalian neurotransmitter/sodium symporters (NSSs), we studied the properties of proteomicelles formed by n-dodecyl-β,D-maltopyranoside (DDM) detergent. Extensive atomistic molecular dynamics simulations of different protein/detergent/water number ratios revealed the formation of a proteomicelle characterized by a constant-sized shell of detergents surrounding LeuT protecting its transmembrane segments from unfavorable hydrophobic/hydrophilic exposure. Regardless of the DDM content in the simulated system, this shell consisted of a constant number of DDM molecules (∼120 measured at a 4 Å cutoff distance from LeuT). In contrast, the overall number of DDMs in the proteomicelle (aggregation number) was found to depend on the detergent concentration, reaching a saturation value of 226±17 DDMs in the highest concentration regime simulated. Remarkably, we found that at high detergent-to-protein ratios we observed two independent ways of DDM penetration into LeuT, both leading to a positioning of the DDM molecule in the second substrate (S2) binding site of LeuT. Consonant with several recent experimental studies demonstrating changes in functional properties of membrane proteins due to detergent, our findings highlight how the environment in which the membrane proteins are examined may affect the outcome and interpretation of their mechanistic features.
authors: Department of Physiology and Biophysics, Weill Cornell Medical College of Cornell University (WCMC) , New York, New York 10065, United States.
authors: George
authors: G
authors: Khelashvili
authors: Michael V
authors: MV
authors: LeVine
authors: Lei
authors: L
authors: Shi
authors: Matthias
authors: M
authors: Quick
authors: Jonathan A
authors: JA
authors: Javitch
authors: Harel
authors: H
authors: Weinstein
mesh_terms: D026905:Amino Acid Transport Systems / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D007930:Leucine / Q000378:metabolism*; D008320:Maltose / Q000031:analogs & derivatives / Q000737:chemistry; D008823:Micelles; D056004:Molecular Dynamics Simulation; D013501:Surface-Active Agents / Q000737:chemistry
chemical_list: D026905:Amino Acid Transport Systems; D001426:Bacterial Proteins; D008823:Micelles; D013501:Surface-Active Agents; C411362:dodecyl maltopyranoside; D008320:Maltose; D007930:Leucine
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