4M1V

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

Summary for 4M1V

• Entry DOI: 10.2210/pdb4m1v/pdb
• Related: 2v3q
• Descriptor: Phosphate-binding protein, PHOSPHATE ION, ACETATE ION, ... (6 entities in total)
• Functional Keywords: phosphate-binding protein, protein binding
• Biological source: unidentified prokaryotic organism
• Cellular location: Secreted: P85173
• Total number of polymer chains: 1
• Total formula weight: 39620.84

Authors

Gonzalez, D.,Hiblot, J.,Darbinian, N.,Miller, J.S.,Gotthard, G.,Amini, S.,Chabriere, E.,Elias, M. (deposition date: 2013-08-04, release date: 2014-01-01, Last modification date: 2024-11-06)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.3 Å)