Residues Leu52 and Leu134 are important for the structural integrity of a nucleotide exchange factor GrpE from Bacillus licheniformis.

Abstract

A DNA fragment encoding Bacillus licheniformis GrpE (BlGrpE) with double mutations at codons 52 and 134 was obtained during PCR cloning. Leu52 and Leu134 in BlGrpE were individually replaced with Pro and His to generate BlGrpE-L52P and BlGrpE-L134H. BlGrpE and BlGrpE-L52P synergistically stimulated the ATPase activity of B. licheniformis DnaK (BlDnaK); however, BlGrpE-L134H and the double-mutated protein (BlGrpE-L52P/L134H) had no co-chaperone function. BlGrpE, BlGrpE-L52P, and BlGrpE-L134H mainly interacted with the monomer of BlDnaK but non-specific interaction was observed for BlGrpE-L52P/L134H. Measurement of intrinsic fluorescence revealed a significant alteration of the microenvironment of aromatic acid residues in the mutant proteins. As compared with BlGrpE, quenching of 208-nm and 222-nm signals were observed in the mutant BlGrpEs and the single-mutated proteins were more sensitive to thermal denaturation. Study holds ProTherm entries: 25225, 25226, 25227 Extra Details: GrpE; Bacillus licheniformis; Site-directed mutagenesis; ATPase activity

Submission Details

ID: agsNZkBc3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:55 p.m.

Version: 1

Publication Details
Liang WC;Lin MG;Chou WM;Chi MC;Chang HP;Lin LL,Int. J. Biol. Macromol. (2009) Residues Leu52 and Leu134 are important for the structural integrity of a nucleotide exchange factor GrpE from Bacillus licheniformis. PMID:19665474
Additional Information

Study Summary

Number of data points 8
Proteins Protein GrpE
Unique complexes 3
Assays/Quantities/Protocols Experimental Assay: Tm ; Experimental Assay: dHvH ; Derived Quantity: dTm
Libraries Mutations for sequence MAEEKQNEELNEQEELNETEAETAEAEQTAAEADAPAEETQTEMLEKQLKELQERLEEKENKLLRVQADFENYKRRARLDLEAAEKYRSQRIISDLLPALDNFERALQIDPDNEQTKSLLQGMEMVHRQILEALKNEGVEQIPSVGEQFDPNMHQAVMQVEDEAYESNAVVEELQKGYKLKDRVIRPSMVKVNQ

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 Protein GrpE Q65H53 GRPE_BACLD