Application of Site-Specific 19F Paramagnetic Relaxation Enhancement to Distinguish two Different Conformations of a Multidomain Protein

被引：5

作者：

Shi, Pan ^{[1
,2
]}

Li, Dong ^{[1
,2
]}

Li, Juan ^{[1
,2
]}

Chen, Hongwei ^{[3
]}

Wu, Fangming ^{[3
]}

Xiong, Ying ^{[1
,2
]}

Tian, Changlin ^{[1
,2
,3
]}

机构：

[1] Univ Sci & Technol China, Hefei Natl Lab Phys Sci Microscale, Hefei 230026, Peoples R China

[2] Univ Sci & Technol China, Sch Life Sci, Hefei 230026, Peoples R China

[3] Chinese Acad Sci, High Field Magnet Lab, Hefei 230031, Peoples R China

来源：

JOURNAL OF PHYSICAL CHEMISTRY LETTERS | 2012年 / 3卷 / 01期

关键词：

NMR; PROBE;

D O I：

10.1021/jz201480g

中图分类号：

O64 [物理化学（理论化学）、化学物理学];

学科分类号：

070304 ; 081704 ;

摘要：

Paramagnetic relaxation enhancement (PRE) provides long-distance restraints in solution NMR protein structural studies. It has been shown previously that L27tan, a protein with tandem L27-domains, has two possible conformations. Here, F-19 was site-specifically introduced to L27tan via the incorporation of an unnatural amino acid, trifluoromethyl-phenylalanine (tfmF). Different F-19 signal intensity attenuations were observed at different L27tan sites, due to different distances between the site-specifically incorporated tfmF and site-directed spin radical labeling. Analysis of the F-19 detection PRE showed that the L27tan protein had a closed conformation in solution. This F-19 detection PRE method could be further applied in distance measurements for proteins of large size, including multidomain proteins or membrane proteins.

引用

页码：34 / 37

页数：4

共 11 条

[1] Utilization of site-directed spin labeling and high-resolution heteronuclear nuclear magnetic resonance for global fold determination of large proteins with limited nuclear overhauser effect data [J].

Battiste, JL ;

Wagner, G .

BIOCHEMISTRY, 2000, 39 (18) :5355-5365

[2] Use of F-19 NMR to probe protein structure and conformational changes [J].

Danielson, MA ;

Falke, JJ .

ANNUAL REVIEW OF BIOPHYSICS AND BIOMOLECULAR STRUCTURE, 1996, 25 :163-195

[3] Characterization of long-range structure in the denatured state of staphylococcal nuclease .1. Paramagnetic relaxation enhancement by nitroxide spin labels [J].

Gillespie, JR ;

Shortle, D .

JOURNAL OF MOLECULAR BIOLOGY, 1997, 268 (01) :158-169

[4] Preparation of site-specifically labeled fluorinated proteins for 19F-NMR structural characterization [J].

Hammill, Jared T. ;

Miyake-Stoner, Shigeki ;

Hazen, Jennifer L. ;

Jackson, Jennifer C. ;

Mehl, Ryan A. .

NATURE PROTOCOLS, 2007, 2 (10) :2601-2607

[5] Site-specific incorporation of a 19F-amino acid into proteins as an NMR probe for characterizing protein structure and reactivity [J].

Jackson, Jennifer C. ;

Hammill, Jared T. ;

Mehl, Ryan A. .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2007, 129 (05) :1160-1166

[6] Structural Analysis of Protein Interfaces from 13C Direct-Detected Paramagnetic Relaxation Enhancements [J].

Madl, Tobias ;

Felli, Isabella C. ;

Bertini, Ivano ;

Sattler, Michael .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2010, 132 (21) :7285-+

[7] Site-specific labelling with a metal chelator for protein-structure refinement [J].

Pintacuda, G ;

Moshref, A ;

Leonchiks, A ;

Sharipo, A ;

Otting, G .

JOURNAL OF BIOMOLECULAR NMR, 2004, 29 (03) :351-361

[8] Site-specific 19F NMR chemical shift and side chain relaxation analysis of a membrane protein labeled with an unnatural amino acid [J].

Shi, Pan ;

Wang, Hu ;

Xi, Zhaoyong ;

Shi, Chaowei ;

Xiong, Ying ;

Tian, Changlin .

PROTEIN SCIENCE, 2011, 20 (01) :224-228

[9] Site-specific protein backbone and side-chain NMR chemical shift and relaxation analysis of human vinexin SH3 domain using a genetically encoded 15N/19F-labeled unnatural amino acid [J].

Shi, Pan ;

Xi, Zhaoyong ;

Wang, Hu ;

Shi, Chaowei ;

Xiong, Ying ;

Tian, Changlin .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2010, 402 (03) :461-466

[10] RELAXATION PROCESSES IN A SYSTEM OF 2 SPINS [J].

SOLOMON, I .

PHYSICAL REVIEW, 1955, 99 (02) :559-565

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