title: MQ-HNCO-TROSY for the measurement of scalar and residual dipolar couplings in larger proteins: application to a 557-residue IgFLNa16-21.
abstract: We describe a novel pulse sequence, MQ-HNCO-TROSY, for the measurement of scalar and residual dipolar couplings between amide proton and nitrogen in larger proteins. The experiment utilizes the whole 2T(N) polarization transfer delay for labeling of (15)N chemical shift in a constant time manner, which efficiently doubles the attainable resolution in (15)N dimension with respect to the conventional HNCO-TROSY experiment. In addition, the accordion principle is employed for measuring (J + D)(NH)s, and the multiplet components are selected with the generalized version of the TROSY scheme introduced by Nietlispach (J Biomol NMR 31:161-166, 2005). Therefore, cross peak overlap is diminished while the time period during which the (15)N spin is susceptible to fast transverse relaxation associated with the anti-TROSY transition is minimized per attainable resolution unit. The proposed MQ-HNCO-TROSY scheme was employed for measuring RDCs in high molecular weight protein IgFLNa16-21 of 557 residues, resulting in 431 experimental RDCs. Correlations between experimental and back-calculated RDCs in individual domains gave relatively low Q-factors (0.19-0.39), indicative of sufficient accuracy that can be obtained with the proposed MQ-HNCO-TROSY experiment in high molecular weight proteins.
authors.affiliation: NMR Laboratory, Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR Laboratory, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland.
authors.forename: Sampo
authors.initials: S
authors.lastname: Mäntylahti
authors.forename: Outi
authors.initials: O
authors.lastname: Koskela
authors.forename: Pengju
authors.initials: P
authors.lastname: Jiang
authors.forename: Perttu
authors.initials: P
authors.lastname: Permi
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D002247:Carbon Isotopes / Q000737:chemistry; D003285:Contractile Proteins / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry; D064448:Filamins; D005583:Fourier Analysis; D006801:Humans; D008840:Microfilament Proteins / Q000737:chemistry; D009587:Nitrogen Isotopes / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D025801:Ubiquitin / Q000737:chemistry; D014867:Water / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D002247:Carbon Isotopes; D003285:Contractile Proteins; D064448:Filamins; C034284:IgG Fc-binding protein, Streptococcus; D008840:Microfilament Proteins; D009587:Nitrogen Isotopes; D011506:Proteins; D025801:Ubiquitin; D014867:Water; D003903:Deuterium
references.citation: J Biomol NMR. 2000 May;17(1):43-54
references.pmid: 10909865
references.citation: Methods Enzymol. 2001;339:127-74
references.pmid: 11462810
references.citation: Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12366-71
references.pmid: 9356455
references.citation: J Biomol NMR. 1999 Jan;13(1):3-10
references.pmid: 21080259
references.citation: J Biomol NMR. 2006 Feb;34(2):101-15
references.pmid: 16518697
references.citation: J Biol Chem. 2009 Sep 11;284(37):25450-8
references.pmid: 19622754
references.citation: Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13585-90
references.pmid: 9811843
references.citation: J Magn Reson. 1999 Mar;137(1):243-6
references.pmid: 10053154
references.citation: J Biomol NMR. 1997 Oct;10(3):301-6
references.pmid: 20700833
references.citation: Nat Struct Biol. 1998 Dec;5(12):1065-74
references.pmid: 9846877
references.citation: Eur J Biochem. 2000 Nov;267(22):6665-72
references.pmid: 11054120
references.citation: J Magn Reson. 2000 Oct;146(2):255-9
references.pmid: 11001841
references.citation: Biochemistry. 1999 Jul 13;38(28):9013-22
references.pmid: 10413474
references.citation: J Mol Biol. 2003 Apr 11;327(5):1121-33
references.pmid: 12662935
references.citation: J Biomol NMR. 2002 Jan;22(1):27-35
references.pmid: 11885978
references.citation: Q Rev Biophys. 2000 Nov;33(4):371-424
references.pmid: 11233409
references.citation: J Magn Reson. 1998 Aug;133(2):364-7
references.pmid: 9716480
references.citation: Chemphyschem. 2007 Sep 17;8(13):1901-9
references.pmid: 17654630
references.citation: J Biomol NMR. 2005 Feb;31(2):161-6
references.pmid: 15772756
references.citation: J Magn Reson. 1999 Jun;138(2):334-42
references.pmid: 10341140
references.citation: J Am Chem Soc. 2004 Oct 6;126(39):12646-50
references.pmid: 15453798
references.citation: EMBO J. 2007 Sep 5;26(17):3993-4004
references.pmid: 17690686
references.citation: Science. 1997 Nov 7;278(5340):1111-4
references.pmid: 9353189
references.citation: J Biomol NMR. 2003 Dec;27(4):341-9
references.pmid: 14512731
references.citation: J Biomol NMR. 1997 Jul;10(1):89-94
references.pmid: 9453800
references.citation: J Biomol NMR. 2010 Feb;46(2):175-89
references.pmid: 20012159
references.citation: J Magn Reson. 2000 Mar;143(1):184-96
references.pmid: 10698659
references.citation: Magn Reson Chem. 2006 Jul;44 Spec No:S168-76
references.pmid: 16823899
references.citation: J Magn Reson. 1999 Sep;140(1):259-63
references.pmid: 10479570
references.citation: J Magn Reson. 2009 Oct;200(2):173-7
references.pmid: 19615926
references.citation: J Magn Reson. 1998 Apr;131(2):373-8
references.pmid: 9571116
references.citation: Chem Rev. 2006 May;106(5):1720-36
references.pmid: 16683751