Thermal stability and cooperative domains of CPL1 lysozyme and its NH2- and COOH-terminal modules. Dependence on choline binding.


Abstract

Differential scanning calorimetry (DSC) has been employed to characterize the thermal denaturation of CPL1 lysozyme and its isolated fragments in the absence and presence of choline. The heat capacity function of CPL1 lysozyme shows two peaks with Tm values of 43.5 and 51.4 degrees C. At saturating concentrations of choline the second transition disappears, and the Tm is shifted to higher temperatures. The DSC thermogram of the C-CPL1 protein corresponding to the carboxyl-terminal domain of CPL1 lysozyme has a single peak with a Tm of 42.9 degrees C. The effect of choline is very similar to that observed for the whole CPL1 lysozyme. The NH2-terminal fragment obtained by proteolytic digestion shows a Tm of 52 degrees C, close to that of 51.4 degrees C found for the second transition of CPL1, and choline does not affect the Tm nor the denaturation enthalpy. These data suggest that choline is bound to the COOH-terminal domain of the protein. Deconvolution of the excess heat capacity curve of the CPL1 lysozyme shows that the data can be fitted to two two-state independent transitions. The analysis of the DSC curves showed that the NH2-terminal unfolding enthalpy steadily decreases with increasing concentrations of choline. These results indicate that, under saturating concentrations of choline, whole CPL1 could unfold as a single cooperative unit. Study holds ProTherm entries: 5144, 5145, 5146 Extra Details: transition 1; C-terminal module; reversibility is from scanning-rate,independent thermogram domain; CPL1 lysozyme; deconvolution;,two-state independent transitions; cooperative unit

Submission Details

ID: 7fwJWmH73

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:28 p.m.

Version: 1

Publication Details
Sanz JM;García JL;Laynez J;Usobiaga P;Menéndez M,J. Biol. Chem. (1993) Thermal stability and cooperative domains of CPL1 lysozyme and its NH2- and COOH-terminal modules. Dependence on choline binding. PMID:8454587
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
2J8G 2007-07-03 1.69 Crystal structure of the modular Cpl-1 endolysin complexed with a peptidoglycan analogue (E94Q mutant in complex with a tetrasaccharide- pentapeptide)
2J8F 2007-07-03 1.84 Crystal structure of the modular Cpl-1 endolysin complexed with a peptidoglycan analogue (E94Q mutant in complex with a disaccharide- pentapeptide)
2IXV 2007-07-03 1.96 Crystal structure of the modular Cpl-1 endolysin complexed with a peptidoglycan analogue (E94Q mutant)
1H09 2003-06-26 2.1 Multimodular Pneumococcal Cell Wall Endolysin from phage Cp-1
2IXU 2007-07-03 2.28 Crystal structure of the modular Cpl-1 endolysin complexed with a peptidoglycan analogue (wild-type endolysin)
1OBA 2003-10-17 2.45 Multimodular Pneumococcal Cell Wall Endolysin from phage Cp-1 complexed with choline

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
97.1 Lysozyme P19386 LYS_BPCP9
100.0 Lysozyme P15057 LYS_BPCP1