Protein alignment by a coexpressed lanthanide-binding tag for the measurement of residual dipolar couplings

被引：160

作者：

Wöhnert, J

Franz, KJ

Nitz, M

Imperiali, B

Schwalbe, H

机构：

[1] MIT, Dept Chem, Cambridge, MA 02139 USA

[2] MIT, Dept Biol, Cambridge, MA 02139 USA

[3] Goethe Univ Frankfurt, Inst Organ Chem & Chem Biol, Ctr Biomol Magnet Resonance, D-60439 Frankfurt, Germany

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2003年 / 125卷 / 44期

关键词：

D O I：

10.1021/ja036022d

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

A protein fusion construct of human ubiquitin with an N-terminal lanthanide binding tag (LBT) enables observation of long-range orientational restraints in solution NMR from residual dipolar couplings (RDCs) due to paramagnetic alignment of the protein. The paramagnetic lanthanide ions Tb3+, Dy3+, and Tm3+ are shown to bind to the LBT and induce different alignment tensors, in agreement with theory. RDCs, measured relative to the diamagnetic Lu3+, range from ?7.6 to 5.5 Hz for Tb3+ and ?6.6 to 6.1 Hz for Dy3+, while an opposite alignment tensor is observed for Tm3+ (4.5 to ?2.9 Hz) at 800 MHz. Experimental RDCs are in excellent agreement with those predicted on the basis of the X-ray structure of the protein. Copyright © 2003 American Chemical Society.

引用

收藏

页码：13338 / 13339

页数：2

相关论文

共 32 条

[1] Lanthanide-induced pseudocontact shifts for solution structure refinements of macromolecules in shells up to 40 Å from the metal ion [J].

Allegrozzi, M ;

Bertini, I ;

Janik, MBL ;

Lee, YM ;

Lin, GH ;

Luchinat, C .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2000, 122 (17) :4154-4161

[2] The auto-orientation in high magnetic fields of oxidized cytochrome b562 as source of constraints for solution structure determination [J].

Arnesano, F ;

Banci, L ;

Bertini, I ;

van der Wetering, K ;

Czisch, M ;

Kaptein, R .

JOURNAL OF BIOMOLECULAR NMR, 2000, 17 (04) :295-304

[3] Partial orientation of oxidized and reduced cytochrome b5 at high magnetic fields:: Magnetic susceptibility anisotropy contributions and consequences for protein solution structure determination [J].

Banci, L ;

Bertini, I ;

Huber, JG ;

Luchinat, C ;

Rosato, A .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1998, 120 (49) :12903-12909

[4] Paramagnetically induced residual dipolar couplings for solution structure determination of lanthanide binding proteins [J].

Barbieri, R ;

Bertini, I ;

Cavallaro, G ;

Lee, YM ;

Luchinat, C ;

Rosato, A .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (19) :5581-5587

[5] Dipolar couplings in macromolecular structure determination [J].

Bax, A ;

Kontaxis, G ;

Tjandra, N .

NUCLEAR MAGNETIC RESONANCE OF BIOLOGICAL MACROMOLECULES, PT B, 2001, 339 :127-174

[6] Magnetic susceptibility tenser anisotropies for a lanthanide ion series in a fixed protein matrix [J].

Bertini, I ;

Janik, MBL ;

Lee, YM ;

Luchinat, C ;

Rosato, A .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2001, 123 (18) :4181-4188

[7] NMR approaches for monitoring domain orientations in calcium-binding proteins in solution using partial replacement of Ca2+ by Tb3+ [J].

Biekofsky, RR ;

Muskett, FW ;

Schmidt, JM ;

Martin, SR ;

Browne, JP ;

Bayley, PM ;

Feeney, J .

FEBS LETTERS, 1999, 460 (03) :519-526

[8] Measurement of residual dipolar couplings of macromolecules aligned in the nematic phase of a colloidal suspension of rod-shaped viruses [J].

Clore, GM ;

Starich, MR ;

Gronenborn, AM .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1998, 120 (40) :10571-10572

[9] Measurement of one bond dipolar couplings through lanthanide-induced orientation of a calcium-binding protein [J].

Contreras, MA ;

Ubach, J ;

Millet, O ;

Rizo, J ;

Pons, M .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1999, 121 (38) :8947-8948

[10] TWO-DIMENSIONAL H-1-NMR STUDY OF HUMAN UBIQUITIN - A MAIN CHAIN DIRECTED ASSIGNMENT AND STRUCTURE-ANALYSIS [J].

DISTEFANO, DL ;

WAND, AJ .

BIOCHEMISTRY, 1987, 26 (23) :7272-7281

← 1 2 3 4 →