title: Probing structure and dynamics of protein assemblies by magic angle spinning NMR spectroscopy.
abstract: In living organisms, biological molecules often organize into multicomponent complexes. Such assemblies consist of various proteins and carry out essential functions, ranging from cell division, transport, and energy transduction to catalysis, signaling, and viral infectivity. To understand the biological functions of these assemblies, in both healthy and disease states, researchers need to study their three-dimensional architecture and molecular dynamics. To date, the large size, the lack of inherent long-range order, and insolubility have made atomic resolution studies of many protein assemblies challenging or impractical using traditional structural biology methods such as X-ray diffraction and solution NMR spectroscopy. In the past 10 years, we have focused our work on the development and application of magic angle spinning solid-state NMR (MAS NMR) methods to characterize large protein assemblies at atomic-level resolution. In this Account, we discuss the rapid progress in the field of MAS NMR spectroscopy, citing work from our laboratory and others on methodological developments that have facilitated the in-depth analysis of biologically important protein assemblies. We emphasize techniques that yield enhanced sensitivity and resolution, such as fast MAS (spinning frequencies of 40 kHz and above) and nonuniform sampling protocols for data acquisition and processing. We also discuss the experiments for gaining distance restraints and for recoupling anisotropic tensorial interactions under fast MAS conditions. We give an overview of sample preparation approaches when working with protein assemblies. Following the overview of contemporary MAS NMR methods, we present case studies into the structure and dynamics of two classes of biological systems under investigation in our laboratory. We will first turn our attention to cytoskeletal microtubule motor proteins including mammalian dynactin and dynein light chain 8. We will then discuss protein assemblies from the HIV-1 retrovirus.
authors: Department of Chemistry and Biochemistry, University of Delaware , Newark, Delaware 19716, United States.
authors: Si
authors: S
authors: Yan
authors: Christopher L
authors: CL
authors: Suiter
authors: Guangjin
authors: G
authors: Hou
authors: Huilan
authors: H
authors: Zhang
authors: Tatyana
authors: T
authors: Polenova
mesh_terms: D000818:Animals; D003599:Cytoskeleton / Q000737:chemistry / Q000378:metabolism; D015497:HIV-1 / Q000737:chemistry / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D019906:Nuclear Magnetic Resonance, Biomolecular; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D015203:Reproducibility of Results; D012191:Retroviridae Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D011506:Proteins; D012191:Retroviridae Proteins
references: J Am Chem Soc. 2007 Jun 20;129(24):7502-3
references: 17530852
references: Can J Chem. 2011;89(7):909-918
references: 23243318
references: J Am Chem Soc. 2004 Dec 22;126(50):16608-20
references: 15600367
references: Magn Reson Chem. 2011 Aug;49(8):483-91
references: 21751244
references: J Phys Chem B. 2012 Jun 28;116(25):7416-27
references: 22667827
references: J Am Chem Soc. 2012 Sep 12;134(36):14730-3
references: 22916960
references: J Am Chem Soc. 2008 Apr 30;130(17):5798-807
references: 18393505
references: Angew Chem Int Ed Engl. 2010 Nov 2;49(45):8346-57
references: 20941715
references: Hum Mol Genet. 2005 Oct 15;14 Spec No. 2:R291-300
references: 16244328
references: Angew Chem Int Ed Engl. 2012 Mar 19;51(12):2998-3001
references: 22298470
references: Nat Methods. 2009 Mar;6(3):215-8
references: 19198596
references: J Am Chem Soc. 2009 Jul 29;131(29):10113-26
references: 19580321
references: J Am Chem Soc. 2009 Sep 30;131(38):13690-702
references: 19736935
references: J Am Chem Soc. 2010 Feb 17;132(6):1976-87
references: 20092249
references: J Chem Phys. 2012 Oct 7;137(13):134201
references: 23039592
references: Proc Natl Acad Sci U S A. 2011 Jun 28;108(26):10396-9
references: 21670262
references: J Magn Reson. 2013 Jul;232:18-30
references: 23685715
references: Magn Reson Chem. 2007 Dec;45 Suppl 1:S84-92
references: 18157839
references: Phys Chem Chem Phys. 2009 Aug 28;11(32):7078-86
references: 19652843
references: J Am Chem Soc. 2007 Feb 28;129(8):2338-44
references: 17279748
references: J Biol Chem. 2008 Oct 3;283(40):27314-24
references: 18650427
references: J Am Chem Soc. 2007 Sep 5;129(35):10650-1
references: 17691789
references: J Am Chem Soc. 2011 Mar 23;133(11):3943-53
references: 21361320
references: J Am Chem Soc. 2009 Nov 11;131(44):15968-9
references: 19839609
references: Curr Opin Struct Biol. 2008 Apr;18(2):203-17
references: 18406133
references: J Phys Chem B. 2012 Nov 26;116(46):13585-96
references: 23094591
references: Proc Natl Acad Sci U S A. 2008 Jul 29;105(30):10366-71
references: 18653759
references: J Am Chem Soc. 2011 Nov 23;133(46):18646-55
references: 21995349
references: J Am Chem Soc. 2004 Apr 21;126(15):4764-5
references: 15080665
references: J Magn Reson. 2009 Feb;196(2):133-41
references: 19028122
references: Angew Chem Int Ed Engl. 2010 Nov 22;49(48):9215-8
references: 20957710
references: Phys Chem Chem Phys. 2010 Dec 7;12(45):14873-83
references: 20936218
references: Protein Sci. 2010 Apr;19(4):716-30
references: 20095046
references: J Am Chem Soc. 2012 Apr 11;134(14):6455-66
references: 22428579
references: Biochemistry. 2010 Jun 29;49(25):5083-5
references: 20518521
references: Cell. 2003 Feb 21;112(4):467-80
references: 12600311
references: Solid State Nucl Magn Reson. 2012 May-Jun;43-44:1-13
references: 22464402
references: J Am Chem Soc. 2006 Aug 9;128(31):9992-3
references: 16881610
references: J Am Chem Soc. 2007 Sep 26;129(38):11791-801
references: 17725352
references: Magn Reson Chem. 2007 Dec;45 Suppl 1:S73-83
references: 18157811