2EYC

DMSO refined solution structure of crambin in dpc micelles

2EYC の概要

• エントリーDOI: 10.2210/pdb2eyc/pdb
• 関連するPDBエントリー: 1YV8 1YVA 2EYA 2EYB 2EYD
• 分子名称: CRAMBIN (1 entity in total)
• 機能のキーワード: crambin, dpc, micelles, plant protein
• 由来する生物種: Crambe hispanica subsp. abyssinica
• 細胞内の位置: Secreted: P01542
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 4738.45

構造登録者

Ahn, H.C.,Markley, J.L. (登録日: 2005-11-09, 公開日: 2006-05-23, 最終更新日: 2024-10-30)

主引用文献

Ahn, H.C.,Juranic, N.,Macura, S.,Markley, J.L.
Three-dimensional structure of the water-insoluble protein crambin in dodecylphosphocholine micelles and its minimal solvent-exposed surface.
J.Am.Chem.Soc., 128:4398-4404, 2006

PubMed Abstract: We chose crambin, a hydrophobic and water-insoluble protein originally isolated from the seeds of the plant Crambe abyssinica, as a model for NMR investigations of membrane-associated proteins. We produced isotopically labeled crambin(P22,L25) (variant of crambin containing Pro22 and Leu25) as a cleavable fusion with staphylococcal nuclease and refolded the protein by an approach that has proved successful for the production of proteins with multiple disulfide bonds. We used NMR spectroscopy to determine the three-dimensional structure of the protein in two membrane-mimetic environments: in a mixed aqueous-organic solvent (75%/25%, acetone/water) and in DPC micelles. With the sample in the mixed solvent, it was possible to determine (>NH...OC<) hydrogen bonds directly by the detection of (h3)J(NC)' couplings. H-bonds determined in this manner were utilized in the refinement of the NMR-derived protein structures. With the protein in DPC (dodecylphosphocholine) micelles, we used manganous ion as an aqueous paramagnetic probe to determine the surface of crambin that is shielded by the detergent. With the exception of the aqueous solvent exposed loop containing residues 20 and 21, the protein surface was protected by DPC. This suggests that the protein may be similarly embedded in physiological membranes. The strategy described here for the expression and structure determination of crambin should be applicable to structural and functional studies of membrane active toxins and small membrane proteins.

PubMed: 16569017
DOI: 10.1021/ja057773d

実験手法

SOLUTION NMR