Structural highlights
Disease
PHBP_UNKP May be involved in atherosclerosis.
Function
PHBP_UNKP Phosphate-binding protein.[1]
Publication Abstract from PubMed
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 A, respectively).
Ancestral mutations as a tool for solubilizing proteins: The case of a hydrophobic phosphate-binding protein.,Gonzalez D, Hiblot J, Darbinian N, Miller JC, Gotthard G, Amini S, Chabriere E, Elias M FEBS Open Bio. 2014 Jan 3;4:121-7. doi: 10.1016/j.fob.2013.12.006. eCollection, 2014. PMID:24490136[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Diemer H, Elias M, Renault F, Rochu D, Contreras-Martel C, Schaeffer C, Van Dorsselaer A, Chabriere E. Tandem use of X-ray crystallography and mass spectrometry to obtain ab initio the complete and exact amino acids sequence of HPBP, a human 38-kDa apolipoprotein. Proteins. 2008 Jun;71(4):1708-20. PMID:18076037 doi:10.1002/prot.21866
- ↑ Gonzalez D, Hiblot J, Darbinian N, Miller JC, 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. 2014 Jan 3;4:121-7. doi: 10.1016/j.fob.2013.12.006. eCollection, 2014. PMID:24490136 doi:http://dx.doi.org/10.1016/j.fob.2013.12.006