Thermodynamic and functional characterization of protein W from bacteriophage lambda. The three C-terminal residues are critical for activity.


Abstract

Gene product W (gpW), the head-tail joining protein from bacteriophage lambda, provides a fascinating model for studying protein interactions. Composed of only 68 residues, it must interact with at least two other proteins in the phage, and probably with DNA. To study the structural and functional properties of gpW, plasmids were constructed expressing gpW with hexahistidine tag sequences at either the N or C terminus. The purified wild type fusion proteins were found to be stably folded and biologically active. The protein is monomeric as judged by equilibrium ultracentrifugation, and appears to unfold by a cooperative two-state mechanism. Circular dichroism studies indicate that the protein is 47% helical, with a T(m) of 71.3 degrees C, and a DeltaG(u) of 3.01 kcal/mol at 25 degrees C. Mutagenesis of the three hydrophobic C-terminal residues of gpW showed that they are critical for activity, even though they do not contribute to the thermodynamic stability of the protein. Using secondary structure prediction as a guide, we also designed destabilized gpW mutants. The hydrophobic nature of the gpW C terminus caused these mutants to be degraded by the ClpP-containing proteases in Escherichia coli. Study holds ProTherm entries: 8511, 8512, 8513, 8514 Extra Details: additive : EDTA(0.2 mM),(C terminus) gene product W; head-tail joining protein; biologically active;,hydrophobic; activity

Submission Details

ID: TARe2g8J

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:36 p.m.

Version: 1

Publication Details
Maxwell KL;Davidson AR;Murialdo H;Gold M,J. Biol. Chem. (2000) Thermodynamic and functional characterization of protein W from bacteriophage lambda. The three C-terminal residues are critical for activity. PMID:10770927
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 PDB Entries

Structure ID Release Date Resolution Structure Title
2L6R 2011-11-30 High resolution NMR structure of gpW (W protein of bacteriophage lambda) at acidic pH
2L6Q 2011-11-16 New high resolution NMR structure of gpW (W protein of bacteriophage lambda) at neutral pH
1HYW 2001-04-25 SOLUTION STRUCTURE OF BACTERIOPHAGE LAMBDA GPW

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Head completion protein P68659 VHTJ_ECOL6
100.0 Head completion protein P68660 HCP_LAMBD