ATP-dependent Conformational Changes Trigger Substrate Capture and Release by an ECF-type Biotin Transporter

Finkenwirth, Friedrich; Sippach, Michael; Landmesser, Heidi; Kirsch, Franziska; Ogienko, Anastasia; Grunzel, Miriam; Kiesler, Cornelia; Steinhoff, Heinz‐Jürgen; Schneider, Erwin; Eitinger, Thomas

doi:10.1074/jbc.m115.654343

Cited by 24 publications

(51 citation statements)

References 71 publications

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“…102 A biotin transporter from Rhodobacter capsulatus was expressed in E.coli , purified and incorporated into Nanodiscs using a total E.coli lipid mixture. In order to successfully incorporate a multi-subunit transporter, the extended scaffold protein MSP1E3D1 was used to generate larger Nanodiscs.…”

Section: Application Of Nanodiscs In Functional Studies Of Membranmentioning

confidence: 99%

Nanodiscs in Membrane Biochemistry and Biophysics

2017

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Membrane proteins play a most important part in metabolism, signaling, cell motility, transport, development, and many other biochemical and biophysical processes which constitute fundamentals of life on molecular level. Detailed understanding of these processes is necessary for the progress of life sciences and biomedical applications. Nanodiscs provide a new and powerful tool for a broad spectrum of biochemical and biophysical studies of membrane proteins and are commonly acknowledged as an optimal membrane mimetic system which provides control over size, composition and specific functional modifications on nanometer scale. In this review we attempted to combine a comprehensive list of various applications of Nanodisc technology with systematic analysis of the most attractive features of this system and advantages provided by Nanodiscs for structural and mechanistic studies of membrane proteins.

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Section: Application Of Nanodiscs In Functional Studies Of Membranmentioning

confidence: 99%

Nanodiscs in Membrane Biochemistry and Biophysics

2017

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“…Sometimes, natural lipid extracts (e.g., Escherichia coli polar or total lipid extract) containing a mixture of lipids are utilized for nanodisc reconstitution, particularly if the membrane protein has been demonstrated to be functionally active in these lipid extracts (Finkenwirth et al., ). In a few cases, protein‐incorporated nanodiscs are assembled directly from cell membranes or native tissue membranes (Gregersen, Fedosova, Nissen, & Boesen, ).…”

Section: Strategic Planningmentioning

confidence: 99%

Preparation of Lipid Nanodiscs with Lipid Mixtures

Atkins

McClary

2019

CP Protein Science

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Lipid nanodiscs provide a native‐like lipid environment for membrane proteins, and they have become a valuable platform for the study of membrane biophysics. A range of biophysical and biochemical analyses are enabled when membrane proteins are captured in lipid nanodiscs. Two parameters that can be controlled when capturing membrane proteins in lipid nanodiscs are the radius, and hence the surface area of the lipid surface, and the composition of the lipid bilayer. Despite their emergence as a versatile tool, most studies with lipid nanodiscs in the literature have focused on nanodiscs of a single radius with a single lipid. In light of the complexity of biological membranes, it is likely that nanodiscs with multiple membrane components would be more sophisticated models for membrane research. It is possible to prepare nanodiscs with more complex lipid mixtures to probe the effects of lipid composition on several aspects of membrane biochemistry. Detailed protocols are described here for the preparation of nanodiscs with mixtures of phospholipids, incorporation of cholesterol, and incorporation of a spectroscopic lipid probe. These protocols provide starting points for the construction of nanodiscs with more physiological membrane compositions or with useful biophysical probes. © 2019 by John Wiley & Sons, Inc. Basic Protocol 1: Assembly of mixed lipid nanodiscs Basic Protocol 2: Assembly of nanodiscs with cholesterol Basic Protocol 3: Incorporation of laurdan into nanodiscs for membrane fluidity measurements

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“…For type 1 and type 2 importers each one common transport model has been proposed, which are supported by several EPR studies [100][101][102][103][104][105][106][107][108][109][110]. First EPR studies on the dynamics of ECF transporters were presented recently [111,112], while the final mechanism is still under debate. Most EPR studies with a particular focus on pulsed methods, however, are available for ABC exporters [113][114][115][116][117][118][119][120][121][122][123][124] but their data are discussed as controversial as the present structures such that no final, common model has been agreed on.…”

Section: Abc Transportersmentioning

confidence: 74%

The Synergetic Effects of Combining Structural Biology and EPR Spectroscopy on Membrane Proteins

Wunnicke

Hänelt

2017

Crystals

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Protein structures as provided by structural biology such as X-ray crystallography, cryo-electron microscopy and NMR spectroscopy are key elements to understand the function of a protein on the molecular level. Nonetheless, they might be error-prone due to crystallization artifacts or, in particular in case of membrane-imbedded proteins, a mostly artificial environment. In this review, we will introduce different EPR spectroscopy methods as powerful tools to complement and validate structural data gaining insights in the dynamics of proteins and protein complexes such that functional cycles can be derived. We will highlight the use of EPR spectroscopy on membrane-embedded proteins and protein complexes ranging from receptors to secondary active transporters as structural information is still limited in this field and the lipid environment is a particular challenge.

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ATP-dependent Conformational Changes Trigger Substrate Capture and Release by an ECF-type Biotin Transporter

Cited by 24 publications

References 71 publications

Nanodiscs in Membrane Biochemistry and Biophysics

Nanodiscs in Membrane Biochemistry and Biophysics

Preparation of Lipid Nanodiscs with Lipid Mixtures

The Synergetic Effects of Combining Structural Biology and EPR Spectroscopy on Membrane Proteins

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