Nanodiscs and solution NMR: preparation, application and challenges

Nanotechnology Reviews

Puthenveetil Robbins¹  Vinogradova Olga²  Nguyen Khiem³ 

[1] Department of Molecular and Cell Biology, CLAS, University of Connecticut at Storrs, Storrs, CT 06269, USA;Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut at Storrs, 69 North Eagleville Rd, Unit 3092, Storrs, CT 06269, USA, Phone: [phone], Fax: [phone];Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut at Storrs, Storrs, CT 06269, USA;

关键词: beta barrel;    integrin;    membrane proteins;    nanodisc;    nanoscale phospholipid bilayers;    saposin-a;    smalp;    solution nmr;    styrene maleic acid;    transmembrane;   

DOI  :  10.1515/ntrev-2016-0076

来源: DOAJ

【 摘 要 】

Nanodiscs provide an excellent system for the structure-function investigation of membrane proteins. Its direct advantage lies in presenting a water-soluble form of an otherwise hydrophobic molecule, making it amenable to a plethora of solution techniques. Nuclear magnetic resonance is one such high-resolution approach that looks at the structure and dynamics of a protein with atomic level precision. Recently, there has been a breakthrough in making nanodiscs more susceptible for structure determination by solution NMR, yet it still remains to become the preferred choice for a membrane mimetic. In this practical review, we provide a general discourse on nanodisc and its application to solution NMR. We also offer potential solutions to remediate the technical challenges associated with nanodisc preparation and the choice of proper experimental set-ups. Along with discussing several structural applications, we demonstrate an alternative use of nanodiscs for functional studies, where we investigated the phosphorylation of a cell surface receptor, integrin. This is the first successful manifestation of observing activated receptor phosphorylation in nanodisc through NMR. We additionally present an on-column method for nanodisc preparation with multiple strategies and discuss the potential use of alternative nanoscale phospholipid bilayer systems like styrene maleic acid lipid disc and saposin-A lipoprotein disc.

【 授权许可】

Unknown