PDBsum entry 4m1v

Protein binding

PDB id: 4m1v

Contents

Protein chain

378 a.a.

Ligands

PO4

ACT

EDO ×6

GOL

Waters ×650

PDB id:

4m1v

Name:

Protein binding

Title:

Crystal structure of the ancestral soluble variant of the human phosphate binding protein (hpbp)

Structure:

Phosphate-binding protein. Chain: a. Synonym: hpbp. Engineered: yes. Mutation: yes

Source:

Unidentified prokaryotic organism. Organism_taxid: 2725. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.30Å R-factor: 0.109 R-free: 0.143

Authors:

D.Gonzalez,J.Hiblot,N.Darbinian,J.S.Miller,G.Gotthard,S.Amini, E.Chabriere,M.Elias

Key ref:

D.Gonzalez et al. (2014). Ancestral mutations as a tool for solubilizing proteins: The case of a hydrophobic phosphate-binding protein. Febs Open Bio, 4, 121-127. PubMed id: 24490136 DOI: 10.1016/j.fob.2013.12.006

Date:

04-Aug-13 Release date: 01-Jan-14

Protein chain

P85173  (PHBP_UNKP) -  Phosphate-binding protein from Unknown prokaryotic organism

Seq: 376 a.a.

Struc: 378 a.a.*

* PDB and UniProt seqs differ at 22 residue positions (black crosses)

DOI no: 10.1016/j.fob.2013.12.006

Febs Open Bio 4:121-127 (2014)

PubMed id: 24490136

Ancestral mutations as a tool for solubilizing proteins: The case of a hydrophobic phosphate-binding protein.

D.Gonzalez, J.Hiblot, N.Darbinian, J.C.Miller, G.Gotthard, S.Amini, E.Chabriere, M.Elias.

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).