TY - JOUR
T1 - Kinetics of Membrane Protein-Detergent Interactions Depend on Protein Electrostatics
AU - Wolfe, Aaron J.
AU - Gugel, Jack F.
AU - Chen, Min
AU - Movileanu, Liviu
N1 - Funding Information:
We thank Yi-Ching Hsueh, Adam Blanden, and Bach Pham for their assistance during the very early stage of these studies. The authors are also grateful to Motahareh Ghahari Larimi and Lauren Mayse for their comments and stimulating discussions. This research was supported by the U.S. National Institutes of Health Grants GM115442 (to M.C.), GM088403 (to L.M.), and GM129429 (to L.M.).
Publisher Copyright:
© 2018 American Chemical Society.
PY - 2018/10/18
Y1 - 2018/10/18
N2 - Interactions of a membrane protein with a detergent micelle represent a fundamental process with practical implications in structural and chemical biology. Quantitative assessment of the kinetics of protein-detergent complex (PDC) interactions has always been challenged by complicated behavior of both membrane proteins and solubilizing detergents in aqueous phase. Here, we show the kinetic reads of the desorption of maltoside-containing detergents from β-barrel membrane proteins. Using steady-state fluorescence polarization (FP) anisotropy measurements, we recorded real-time, specific signatures of the PDC interactions. The results of these measurements were used to infer the model-dependent rate constants of association and dissociation of the proteomicelles. Remarkably, the kinetics of the PDC interactions depend on the overall protein charge despite the nonionic nature of the detergent monomers. In the future, this approach might be employed for high-throughput screening of kinetic fingerprints of different membrane proteins stabilized in micelles that contain mixtures of various detergents.
AB - Interactions of a membrane protein with a detergent micelle represent a fundamental process with practical implications in structural and chemical biology. Quantitative assessment of the kinetics of protein-detergent complex (PDC) interactions has always been challenged by complicated behavior of both membrane proteins and solubilizing detergents in aqueous phase. Here, we show the kinetic reads of the desorption of maltoside-containing detergents from β-barrel membrane proteins. Using steady-state fluorescence polarization (FP) anisotropy measurements, we recorded real-time, specific signatures of the PDC interactions. The results of these measurements were used to infer the model-dependent rate constants of association and dissociation of the proteomicelles. Remarkably, the kinetics of the PDC interactions depend on the overall protein charge despite the nonionic nature of the detergent monomers. In the future, this approach might be employed for high-throughput screening of kinetic fingerprints of different membrane proteins stabilized in micelles that contain mixtures of various detergents.
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U2 - 10.1021/acs.jpcb.8b07889
DO - 10.1021/acs.jpcb.8b07889
M3 - Article
C2 - 30251852
AN - SCOPUS:85054688598
SN - 1520-6106
VL - 122
SP - 9471
EP - 9481
JO - Journal of Physical Chemistry B
JF - Journal of Physical Chemistry B
IS - 41
ER -