2KOM

Solution structure of humar Par-3b PDZ2 (residues 451-549)

2KOM の概要

• エントリーDOI: 10.2210/pdb2kom/pdb
• NMR情報: BMRB: 16520
• 分子名称: Partitioning defective 3 homolog (1 entity in total)
• 機能のキーワード: par-3b, pdz domain, psi, structural genomics, alternative splicing, cell cycle, cell division, cell junction, cell membrane, coiled coil, cytoplasm, cytoskeleton, membrane, phosphoprotein, polymorphism, tight junction, signaling protein, protein structure initiative, center for eukaryotic structural genomics, cesg
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Endomembrane system: Q8TEW0
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 13394.24

構造登録者

Volkman, B.F.,Tyler, R.C.,Peterson, F.C.,Center for Eukaryotic Structural Genomics (CESG) (登録日: 2009-09-24, 公開日: 2009-11-10, 最終更新日: 2024-05-22)

主引用文献

Jensen, D.R.,Woytovich, C.,Li, M.,Duvnjak, P.,Cassidy, M.S.,Frederick, R.O.,Bergeman, L.F.,Peterson, F.C.,Volkman, B.F.
Rapid, robotic, small-scale protein production for NMR screening and structure determination.
Protein Sci., 19:570-578, 2010

PubMed Abstract: Three-dimensional protein structure determination is a costly process due in part to the low success rate within groups of potential targets. Conventional validation methods eliminate the vast majority of proteins from further consideration through a time-consuming succession of screens for expression, solubility, purification, and folding. False negatives at each stage incur unwarranted reductions in the overall success rate. We developed a semi-automated protocol for isotopically-labeled protein production using the Maxwell-16, a commercially available bench top robot, that allows for single-step target screening by 2D NMR. In the span of a week, one person can express, purify, and screen 48 different (15)N-labeled proteins, accelerating the validation process by more than 10-fold. The yield from a single channel of the Maxwell-16 is sufficient for acquisition of a high-quality 2D (1)H-(15)N-HSQC spectrum using a 3-mm sample cell and 5-mm cryogenic NMR probe. Maxwell-16 screening of a control group of proteins reproduced previous validation results from conventional small-scale expression screening and large-scale production approaches currently employed by our structural genomics pipeline. Analysis of 18 new protein constructs identified two potential structure targets that included the second PDZ domain of human Par-3. To further demonstrate the broad utility of this production strategy, we solved the PDZ2 NMR structure using [U-(15)N,(13)C] protein prepared using the Maxwell-16. This novel semi-automated protein production protocol reduces the time and cost associated with NMR structure determination by eliminating unnecessary screening and scale-up steps.

PubMed: 20073081
DOI: 10.1002/pro.335

実験手法

SOLUTION NMR