PDBsum entry 2kq4

Immune system

PDB id: 2kq4

Contents

Protein chain

56 a.a. *

* Residue conservation analysis

PDB id:

2kq4

Name:

Immune system

Title:

Atomic resolution protein structure determination by three-dimensional transferred echo double resonance solid-state nuclear magnetic resonance spectroscopy

Structure:

Immunoglobulin g-binding protein g. Chain: x. Fragment: unp residues 303 to 357. Synonym: igg-binding protein g. Engineered: yes. Mutation: yes

Source:

Streptococcus sp. 'Group g'. Organism_taxid: 1320. Gene: spg. Expressed in: escherichia coli. Expression_system_taxid: 562.

NMR struc:

10 models

Authors:

A.J.Nieuwkoop,B.J.Wylie,W.Franks,G.J.Shah,C.M.Rienstra

Key ref:

A.J.Nieuwkoop et al. (2009). Atomic resolution protein structure determination by three-dimensional transferred echo double resonance solid-state nuclear magnetic resonance spectroscopy. J Chem Phys, 131, 095101-095101. PubMed id: 19739873

Date:

27-Oct-09 Release date: 17-Nov-09

Protein chain

P19909  (SPG2_STRSG) -  Immunoglobulin G-binding protein G from Streptococcus sp. group G

Seq: 593 a.a.

Struc: 56 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

J Chem Phys 131:095101-095101 (2009)

PubMed id: 19739873

Atomic resolution protein structure determination by three-dimensional transferred echo double resonance solid-state nuclear magnetic resonance spectroscopy.

A.J.Nieuwkoop, B.J.Wylie, W.T.Franks, G.J.Shah, C.M.Rienstra.

ABSTRACT

We show that quantitative internuclear (15)N-(13)C distances can be obtained in sufficient quantity to determine a complete, high-resolution structure of a moderately sized protein by magic-angle spinning solid-state NMR spectroscopy. The three-dimensional ZF-TEDOR pulse sequence is employed in combination with sparse labeling of (13)C sites in the beta1 domain of the immunoglobulin binding protein G (GB1), as obtained by bacterial expression with 1,3-(13)C or 2-(13)C-glycerol as the (13)C source. Quantitative dipolar trajectories are extracted from two-dimensional (15)N-(13)C planes, in which approximately 750 cross peaks are resolved. The experimental data are fit to exact theoretical trajectories for spin clusters (consisting of one (13)C and several (15)N each), yielding quantitative precision as good as 0.1 A for approximately 350 sites, better than 0.3 A for another 150, and approximately 1.0 A for 150 distances in the range of 5-8 A. Along with isotropic chemical shift-based (TALOS) dihedral angle restraints, the distance restraints are incorporated into simulated annealing calculations to yield a highly precise structure (backbone RMSD of 0.25+/-0.09 A), which also demonstrates excellent agreement with the most closely related crystal structure of GB1 (2QMT, bbRMSD 0.79+/-0.03 A). Moreover, side chain heavy atoms are well restrained (0.76+/-0.06 A total heavy atom RMSD). These results demonstrate for the first time that quantitative internuclear distances can be measured throughout an entire solid protein to yield an atomic-resolution structure.