Computer aided protein engineering to enhance the thermo-stability of CXCR1- T4 lysozyme complex.


Abstract

CXCR1, a member in G-protein coupled receptor (GPCR) family, binds to chemokine interleukin-8 (IL-8) specifically and transduces signals to mediate immune and inflammatory responses. Despite the importance of CXCR1, high-resolution structure determination is hindered by the challenges in crystallization. It has been shown that properly designed mutants with enhanced thermostability, together with fusion partner proteins, can be useful to form crystals for GPCR proteins. In this study, in silico protein design was carried out by using homology modeling and molecular dynamics simulations. To validate the computational modeling results, the thermostability of several mutants and the wild type were measured experimentally. Both computational results and experimental data suggest that the mutant L126W has a significant improvement in the thermostability. This study demonstrated that in silico design can guide protein engineering and potentially facilitate protein crystallography research.

Submission Details

ID: B2h8YFgQ

Submitter: Paul Chang

Submission Date: June 5, 2019, 1:43 p.m.

Version: 1

Publication Details
Wang Y;Park JH;Lupala CS;Yun JH;Jin Z;Huang L;Li X;Tang L;Lee W;Liu H,Sci Rep (2019) Computer aided protein engineering to enhance the thermo-stability of CXCR1- T4 lysozyme complex. PMID:30926935
Additional Information

Study Summary

Number of data points 10
Proteins C-X-C chemokine receptor type 1
Unique complexes 5
Assays/Quantities/Protocols Experimental Assay: Tm ; Experimental Assay: Expression
Libraries Thermostability and expression

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 PDB Entries

Structure ID Release Date Resolution Structure Title
2LNL 2012-10-17 Structure of human CXCR1 in phospholipid bilayers
1ILP 1998-12-23 CXCR-1 N-TERMINAL PEPTIDE BOUND TO INTERLEUKIN-8
1ILQ 1998-12-23 CXCR-1 N-TERMINAL PEPTIDE BOUND TO INTERLEUKIN-8 (MINIMIZED MEAN)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
97.1 C-X-C chemokine receptor type 1 P55919 CXCR1_GORGO
91.9 C-X-C chemokine receptor type 1 Q2YEG0 CXCR1_MACMU
93.5 C-X-C chemokine receptor type 1 Q2YEG2 CXCR1_HOOHO
100.0 C-X-C chemokine receptor type 1 P25024 CXCR1_HUMAN
94.8 C-X-C chemokine receptor type 1 Q2YEF9 CXCR1_PONPY
98.7 C-X-C chemokine receptor type 1 P55920 CXCR1_PANTR