Mapping of unfolding states of integral helical membrane proteins by GPS-NMR and scattering techniques

TFE-induced unfolding of KcsA in DDM surfactant

Antonello Calcutta, Christian Moestrup Jessen, Manja Annette Behrens, Cristiano Luis Pinto De Oliveira, Maria Lourdes Renart, José M González-Ros, Daniel Otzen, Jan Skov Pedersen, Anders Malmendal, Niels Chr Nielsen

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Membrane proteins are vital for biological function, and their action is governed by structural properties critically depending on their interactions with the membranes. This has motivated considerable interest in studies of membrane protein folding and unfolding. Here the structural changes induced by unfolding of an integral membrane protein, namely TFE-induced unfolding of KcsA solubilized by the n-dodecyl ß-d-maltoside (DDM) surfactant is investigated by the recently introduced GPS-NMR (Global Protein folding State mapping by multivariate NMR) (Malmendal et al., PlosONE 5, e10262 (2010)) along with dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS). GPS-NMR is used as a tool for fast analysis of the protein unfolding processes upon external perturbation, and DLS and SAXS are used for further structural characterization of the unfolding states. The combination allows addressing detergent properties and protein conformations at the same time. The mapping of the states reveals that KcsA undergoes a series of rearrangements which include expansion of the tetramer in several steps followed by dissociation into monomers at 29% TFE. Supplementary studies of DDM and TFE in the absence of KcsA suggest that the disintegration of the tetramer at 29% TFE is caused by TFE dissolving the surrounding DDM rim. Above 34% TFE, KcsA collapses to a new structure that is fully formed at 44% TFE.
OriginalsprogEngelsk
TidsskriftBBA General Subjects
Vol/bind181
Udgave nummer9
ISSN0304-4165
DOI
StatusUdgivet - 2012
Udgivet eksterntJa

Citer dette

Calcutta, A., Jessen, C. M., Behrens, M. A., Oliveira, C. L. P. D., Renart, M. L., González-Ros, J. M., ... Nielsen, N. C. (2012). Mapping of unfolding states of integral helical membrane proteins by GPS-NMR and scattering techniques: TFE-induced unfolding of KcsA in DDM surfactant. BBA General Subjects, 181(9). https://doi.org/10.1016/j.bbamem.2012.04.005
Calcutta, Antonello ; Jessen, Christian Moestrup ; Behrens, Manja Annette ; Oliveira, Cristiano Luis Pinto De ; Renart, Maria Lourdes ; González-Ros, José M ; Otzen, Daniel ; Pedersen, Jan Skov ; Malmendal, Anders ; Nielsen, Niels Chr. / Mapping of unfolding states of integral helical membrane proteins by GPS-NMR and scattering techniques : TFE-induced unfolding of KcsA in DDM surfactant. I: BBA General Subjects. 2012 ; Bind 181, Nr. 9.
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title = "Mapping of unfolding states of integral helical membrane proteins by GPS-NMR and scattering techniques: TFE-induced unfolding of KcsA in DDM surfactant",
abstract = "Membrane proteins are vital for biological function, and their action is governed by structural properties critically depending on their interactions with the membranes. This has motivated considerable interest in studies of membrane protein folding and unfolding. Here the structural changes induced by unfolding of an integral membrane protein, namely TFE-induced unfolding of KcsA solubilized by the n-dodecyl {\ss}-d-maltoside (DDM) surfactant is investigated by the recently introduced GPS-NMR (Global Protein folding State mapping by multivariate NMR) (Malmendal et al., PlosONE 5, e10262 (2010)) along with dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS). GPS-NMR is used as a tool for fast analysis of the protein unfolding processes upon external perturbation, and DLS and SAXS are used for further structural characterization of the unfolding states. The combination allows addressing detergent properties and protein conformations at the same time. The mapping of the states reveals that KcsA undergoes a series of rearrangements which include expansion of the tetramer in several steps followed by dissociation into monomers at 29{\%} TFE. Supplementary studies of DDM and TFE in the absence of KcsA suggest that the disintegration of the tetramer at 29{\%} TFE is caused by TFE dissolving the surrounding DDM rim. Above 34{\%} TFE, KcsA collapses to a new structure that is fully formed at 44{\%} TFE.",
author = "Antonello Calcutta and Jessen, {Christian Moestrup} and Behrens, {Manja Annette} and Oliveira, {Cristiano Luis Pinto De} and Renart, {Maria Lourdes} and Gonz{\'a}lez-Ros, {Jos{\'e} M} and Daniel Otzen and Pedersen, {Jan Skov} and Anders Malmendal and Nielsen, {Niels Chr}",
note = "Copyright {\circledC} 2012. Published by Elsevier B.V.",
year = "2012",
doi = "10.1016/j.bbamem.2012.04.005",
language = "English",
volume = "181",
journal = "B B A - General Subjects",
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Mapping of unfolding states of integral helical membrane proteins by GPS-NMR and scattering techniques : TFE-induced unfolding of KcsA in DDM surfactant. / Calcutta, Antonello; Jessen, Christian Moestrup; Behrens, Manja Annette; Oliveira, Cristiano Luis Pinto De; Renart, Maria Lourdes; González-Ros, José M; Otzen, Daniel; Pedersen, Jan Skov; Malmendal, Anders; Nielsen, Niels Chr.

I: BBA General Subjects, Bind 181, Nr. 9, 2012.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Mapping of unfolding states of integral helical membrane proteins by GPS-NMR and scattering techniques

T2 - TFE-induced unfolding of KcsA in DDM surfactant

AU - Calcutta, Antonello

AU - Jessen, Christian Moestrup

AU - Behrens, Manja Annette

AU - Oliveira, Cristiano Luis Pinto De

AU - Renart, Maria Lourdes

AU - González-Ros, José M

AU - Otzen, Daniel

AU - Pedersen, Jan Skov

AU - Malmendal, Anders

AU - Nielsen, Niels Chr

N1 - Copyright © 2012. Published by Elsevier B.V.

PY - 2012

Y1 - 2012

N2 - Membrane proteins are vital for biological function, and their action is governed by structural properties critically depending on their interactions with the membranes. This has motivated considerable interest in studies of membrane protein folding and unfolding. Here the structural changes induced by unfolding of an integral membrane protein, namely TFE-induced unfolding of KcsA solubilized by the n-dodecyl ß-d-maltoside (DDM) surfactant is investigated by the recently introduced GPS-NMR (Global Protein folding State mapping by multivariate NMR) (Malmendal et al., PlosONE 5, e10262 (2010)) along with dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS). GPS-NMR is used as a tool for fast analysis of the protein unfolding processes upon external perturbation, and DLS and SAXS are used for further structural characterization of the unfolding states. The combination allows addressing detergent properties and protein conformations at the same time. The mapping of the states reveals that KcsA undergoes a series of rearrangements which include expansion of the tetramer in several steps followed by dissociation into monomers at 29% TFE. Supplementary studies of DDM and TFE in the absence of KcsA suggest that the disintegration of the tetramer at 29% TFE is caused by TFE dissolving the surrounding DDM rim. Above 34% TFE, KcsA collapses to a new structure that is fully formed at 44% TFE.

AB - Membrane proteins are vital for biological function, and their action is governed by structural properties critically depending on their interactions with the membranes. This has motivated considerable interest in studies of membrane protein folding and unfolding. Here the structural changes induced by unfolding of an integral membrane protein, namely TFE-induced unfolding of KcsA solubilized by the n-dodecyl ß-d-maltoside (DDM) surfactant is investigated by the recently introduced GPS-NMR (Global Protein folding State mapping by multivariate NMR) (Malmendal et al., PlosONE 5, e10262 (2010)) along with dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS). GPS-NMR is used as a tool for fast analysis of the protein unfolding processes upon external perturbation, and DLS and SAXS are used for further structural characterization of the unfolding states. The combination allows addressing detergent properties and protein conformations at the same time. The mapping of the states reveals that KcsA undergoes a series of rearrangements which include expansion of the tetramer in several steps followed by dissociation into monomers at 29% TFE. Supplementary studies of DDM and TFE in the absence of KcsA suggest that the disintegration of the tetramer at 29% TFE is caused by TFE dissolving the surrounding DDM rim. Above 34% TFE, KcsA collapses to a new structure that is fully formed at 44% TFE.

U2 - 10.1016/j.bbamem.2012.04.005

DO - 10.1016/j.bbamem.2012.04.005

M3 - Journal article

VL - 181

JO - B B A - General Subjects

JF - B B A - General Subjects

SN - 0304-4165

IS - 9

ER -