Redesign of LAOBP to bind novel l-amino acid ligands.


Computational protein design is still a challenge for advancing structure-function relationships. While recent advances in this field are promising, more information for genuine predictions is needed. Here, we discuss different approaches applied to install novel glutamine (Gln) binding into the Lysine/Arginine/Ornithine binding protein (LAOBP) from Salmonella typhimurium. We studied the ligand binding behavior of two mutants: a binding pocket grafting design based on a structural superposition of LAOBP to the Gln binding protein QBP from Escherichia coli and a design based on statistical coupled positions. The latter showed the ability to bind Gln even though the protein was not very stable. Comparison of both approaches highlighted a nonconservative shared point mutation between LAOBP_graft and LAOBP_sca. This context dependent L117K mutation in LAOBP turned out to be sufficient for introducing Gln binding, as confirmed by different experimental techniques. Moreover, the crystal structure of LAOBP_L117K in complex with its ligand is reported.

Submission Details

ID: TZNnHzq24

Submitter: Jesus Banda-Vazquez

Submission Date: May 5, 2020, 10:44 a.m.

Version: 1

Publication Details
Banda-Vázquez J;Shanmugaratnam S;Rodríguez-Sotres R;Torres-Larios A;Höcker B;Sosa-Peinado A,Protein Sci (2018) Redesign of LAOBP to bind novel l-amino acid ligands. PMID:29524280
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Lysine/arginine/ornithine-binding periplasmic protein P02911 ARGT_SALTY
93.3 Lysine/arginine/ornithine-binding periplasmic protein P09551 ARGT_ECOLI