Residual methyl protonation in perdeuterated proteins for multi-dimensional correlation experiments in MAS solid-state NMR spectroscopy

被引：65

作者：

Agarwal, Vipin ^{[1
]}

Reif, Bernd ^{[1
,2
]}

机构：

[1] Leibniz Forsch Inst Mol Pharmakol FMP, D-13125 Berlin, Germany

[2] Charite, D-10115 Berlin, Germany

来源：

JOURNAL OF MAGNETIC RESONANCE | 2008年 / 194卷 / 01期

关键词：

MAS solid-state NMR; magic angle spinning; perdeuteration; microcrystalline proteins; TOBSY; TOCSY;

D O I：

10.1016/j.jmr.2008.05.021

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

NMR studies involving perdeuterated proteins focus in general on exchangeable amide protons. However, non-exchangeable sites contain as well a small amount of protons as the employed precursors for protein biosynthesis are not completely proton depleted. The degree of methyl group protonation is in the order of 9% for CD2H using >97% deuterium enriched glucose. We show in this manuscript that this small amount of residual protonation is Sufficient to perform 2D and 3D MAS solid-state NMR experiments. In particular, we suggest a HCCH-TOBSY type experiment which we successfully employ to assign the methyl resonances in aliphatic side chains in a perdeuterated sample of the SH3 domain of chicken alpha-spectrin. (C) 2008 Elsevier Inc. All rights reserved.

引用

页码：16 / 24

页数：9

共 48 条

[41] Ile, Leu, and Val methyl assignments of the 723-residue malate synthase G using a new labeling strategy and novel NMR methods [J].

Tugarinov, V ;

Kay, LE .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (45) :13868-13878

[42] Side chain assignments of Ile δ1 methyl groups in high molecular weight proteins:: An application to a 46 ns tumbling molecule [J].

Tugarinov, V ;

Kay, LE .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (19) :5701-5706

[43] FIXED-POINT THEORY OF ITERATIVE EXCITATION SCHEMES IN NMR [J].

TYCKO, R ;

PINES, A ;

GUCKENHEIMER, J .

JOURNAL OF CHEMICAL PHYSICS, 1985, 83 (06) :2775-2802

[44] High-resolution proton solid-state NMR spectroscopy by phase-modulated Lee-Goldburg experiment [J].

Vinogradov, E ;

Madhu, PK ;

Vega, S .

CHEMICAL PHYSICS LETTERS, 1999, 314 (5-6) :443-450

[45] Proton spectroscopy in solid state nuclear magnetic resonance with windowed phase modulated Lee-Goldburg decoupling sequences [J].

Vinogradov, E ;

Madhu, PK ;

Vega, S .

CHEMICAL PHYSICS LETTERS, 2002, 354 (3-4) :193-202

[46] C-13 CHEMICAL-SHIFT ANISOTROPIES OF SOLID AMINO-ACIDS [J].

YE, CH ;

FU, RQ ;

HU, JZ ;

HOU, L ;

DING, SW .

MAGNETIC RESONANCE IN CHEMISTRY, 1993, 31 (08) :699-704

[47] Solid-State Protein-Structure Determination with Proton-Detected Triple-Resonance 3D Magic-Angle-Spinning NMR Spectroscopy [J].

Zhou, Donghua H. ;

Shea, John J. ;

Nieuwkoop, Andrew J. ;

Franks, W. Trent ;

Wylie, Benjamin J. ;

Mullen, Charles ;

Sandoz, Dennis ;

Rienstra, Chad M. .

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2007, 46 (44) :8380-8383

[48] Proton-detected solid-state NMR Spectroscopy of fully protonated proteins at 40 kHz magic-angle spinning [J].

Zhou, Donghua H. ;

Shah, Gautarn ;

Cormos, Mircea ;

Mullen, Charles ;

Sandoz, Dennis ;

Rienstra, Chad M. .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2007, 129 (38) :11791-11801

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