2GI9

Backbone Conformational Constraints in a Microcrystalline U-15N-Labeled Protein by 3D Dipolar-Shift Solid-State NMR Spectroscopy

Summary for 2GI9

• Entry DOI: 10.2210/pdb2gi9/pdb
• Related: 1PGA
• Descriptor: Immunoglobulin B1 binding domain of protein G (2 entities in total)
• Functional Keywords: gb1, immune system, protein binding
• Biological source: Staphylococcus aureus
• Cellular location: Secreted, cell wall; Peptidoglycan-anchor (Potential): P19909
• Total number of polymer chains: 1
• Total formula weight: 6228.81

Authors

Franks, W.T.,Wylie, B.J.,Stellfox, S.A.,Rienstra, C.M. (deposition date: 2006-03-28, release date: 2006-04-25, Last modification date: 2024-02-14)

Primary citation

Franks, W.T.,Wylie, B.J.,Stellfox, S.A.,Rienstra, C.M.
Backbone Conformational Constraints in a Microcrystalline U-15N-Labeled Protein by 3D Dipolar-Shift Solid-State NMR Spectroscopy
J.Am.Chem.Soc., 128:3154-3155, 2006

PubMed Abstract: Structural studies of uniformly labeled proteins by magic-angle spinning NMR spectroscopy have rapidly matured in recent years. Site-specific chemical shifts of several proteins have been assigned and structures determined from 2D or 3D data sets containing internuclear distance information. Here we demonstrate the application of a complementary technique for constraining protein backbone geometry using a site-resolved 3D dipolar-shift pulse sequence. The dipolar line shapes report on the relative orientations of 1H-15N[i] to 1H-15N[i+1] dipole vectors, constraining the torsion angles phi[i] and psi[i]. In addition, from the same 3D data set, several 1H-15N[i] to1H-15N[i+2] line shapes are extracted to constrain the torsion angles phi[i], psi[i], phi[i+1], and psi[i+1]. We report results for the majority of sites in the 56-residue beta1 immunoglobulin binding domain of protein G (GB1), using 3D experiments at 600 MHz 1H frequency. Excellent agreement between the SSNMR results and a new 1.14 A crystal structure illustrate the general potential of this technique for high-resolution structural refinement of solid proteins.

PubMed: 16522090
DOI: 10.1021/ja058292x

Experimental method

X-RAY DIFFRACTION (1.14 Å)