4M1V

Crystal structure of the ancestral soluble variant of the Human Phosphate Binding Protein (HPBP)

4M1V の概要

• エントリーDOI: 10.2210/pdb4m1v/pdb
• 関連するPDBエントリー: 2v3q
• 分子名称: Phosphate-binding protein, PHOSPHATE ION, ACETATE ION, ... (6 entities in total)
• 機能のキーワード: phosphate-binding protein, protein binding
• 由来する生物種: unidentified prokaryotic organism
• 細胞内の位置: Secreted: P85173
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 39620.84

構造登録者

Gonzalez, D.,Hiblot, J.,Darbinian, N.,Miller, J.S.,Gotthard, G.,Amini, S.,Chabriere, E.,Elias, M. (登録日: 2013-08-04, 公開日: 2014-01-01, 最終更新日: 2024-11-06)

主引用文献

Gonzalez, D.,Hiblot, J.,Darbinian, N.,Miller, J.C.,Gotthard, G.,Amini, S.,Chabriere, E.,Elias, M.
Ancestral mutations as a tool for solubilizing proteins: The case of a hydrophobic phosphate-binding protein.
FEBS Open Bio, 4:121-127, 2014

PubMed Abstract: Stable and soluble proteins are ideal candidates for functional and structural studies. Unfortunately, some proteins or enzymes can be difficult to isolate, being sometimes poorly expressed in heterologous systems, insoluble and/or unstable. Numerous methods have been developed to address these issues, from the screening of various expression systems to the modification of the target protein itself. Here we use a hydrophobic, aggregation-prone, phosphate-binding protein (HPBP) as a case study. We describe a simple and fast method that selectively uses ancestral mutations to generate a soluble, stable and functional variant of the target protein, here named sHPBP. This variant is highly expressed in Escherichia coli, is easily purified and its structure was solved at much higher resolution than its wild-type progenitor (1.3 versus 1.9 Å, respectively).

PubMed: 24490136
DOI: 10.1016/j.fob.2013.12.006

実験手法

X-RAY DIFFRACTION (1.3 Å)