Conformational stability of the Escherichia coli HPr protein: test of the linear extrapolation method and a thermodynamic characterization of cold denaturation.


Abstract

The conformational stability of the histidine-containing phosphocarrier protein (HPr) from Escherichia coli has been determined using a combination of thermal unfolding and urea denaturation experiments. The analysis of the denaturation data provides a measure of the changes in conformational free energy, enthalpy, entropy, and heat capacity that accompany the equilibrium folding of HPr over a wide range of temperature and urea concentrations. In moderate concentrations of urea, HPr undergoes both high- and low-temperature unfolding, allowing for a reliable determination of the change in heat capacity for the conformational transition. The data are consistent with the linear free energy relationship commonly employed to analyze protein denaturation data, even over a relatively large temperature and urea concentration range. Furthermore, we find that a temperature-independent delta Cp is adequate to describe HPr stability over the accessible temperature range. Finally, our data allow us to evaluate the energetics of the urea-protein interaction. For HPr, the changes in excess enthalpy and entropy of the denaturant-protein interaction(s) make only minor contributions to the observed delta H and delta S terms, presumably due in some part to the small size of the HPr protein. Study holds ProTherm entries: 4829, 4830, 4831, 4832, 4833, 4834, 4835, 4836, 4837, 4838, 4839 Extra Details: HPr protein; conformational free energy; size;,temperature-independent delta Cp

Submission Details

ID: ZyNazAXE

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:27 p.m.

Version: 1

Publication Details
Nicholson EM;Scholtz JM,Biochemistry (1996) Conformational stability of the Escherichia coli HPr protein: test of the linear extrapolation method and a thermodynamic characterization of cold denaturation. PMID:8784192
Additional Information

Study Summary

Number of data points 33
Proteins Phosphocarrier protein HPr ; Phosphocarrier protein HPr
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: Cm temp:50.0 C ; Experimental Assay: m temp:50.0 C ; Experimental Assay: dG_H2O temp:50.0 C ; Experimental Assay: Cm temp:45.0 C ; Experimental Assay: m temp:45.0 C ; Experimental Assay: dG_H2O temp:45.0 C ; Experimental Assay: Cm temp:40.0 C ; Experimental Assay: m temp:40.0 C ; Experimental Assay: dG_H2O temp:40.0 C ; Experimental Assay: Cm temp:35.0 C ; Experimental Assay: m temp:35.0 C ; Experimental Assay: dG_H2O temp:35.0 C ; Experimental Assay: Cm temp:30.0 C ; Experimental Assay: m temp:30.0 C ; Experimental Assay: dG_H2O temp:30.0 C ; Experimental Assay: Cm temp:25.0 C ; Experimental Assay: m temp:25.0 C ; Experimental Assay: dG_H2O temp:25.0 C ; Experimental Assay: Cm temp:20.0 C ; Experimental Assay: m temp:20.0 C ; Experimental Assay: dG_H2O temp:20.0 C ; Experimental Assay: Cm temp:15.0 C ; Experimental Assay: m temp:15.0 C ; Experimental Assay: dG_H2O temp:15.0 C ; Experimental Assay: Cm temp:10.0 C ; Experimental Assay: m temp:10.0 C ; Experimental Assay: dG_H2O temp:10.0 C ; Experimental Assay: Cm temp:5.0 C ; Experimental Assay: m temp:5.0 C ; Experimental Assay: dG_H2O temp:5.0 C ; Experimental Assay: Cm temp:0.0 C ; Experimental Assay: m temp:0.0 C ; Experimental Assay: dG_H2O temp:0.0 C
Libraries Mutations for sequence MFQQEVTITAPNGLHTRPAAQFVKEAKGFTSEITVTSNGKSASAKSLFKLQTLGLTQGTVVTISAEGEDEQKAVEHLVKLMAELE

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
2XDF 2010-09-22 Solution Structure of the Enzyme I Dimer Complexed with HPr Using Residual Dipolar Couplings and Small Angle X-Ray Scattering
3EZE 1998-12-16 COMPLEX OF THE AMINO TERMINAL DOMAIN OF ENZYME I AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
1GGR 2000-11-15 COMPLEX OF ENZYME IIAGLC AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
2LRL 2012-05-16 Solution Structures of the IIA(Chitobiose)-HPr complex of the N,N'-Diacetylchitobiose Branch of the Escherichia coli Phosphotransferase System
1HDN 1994-06-22 THE HIGH-RESOLUTION STRUCTURE OF THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI DETERMINED BY RESTRAINED MOLECULAR DYNAMICS FROM NMR NUCLEAR OVERHAUSER EFFECT DATA
1VRC 2005-04-19 Complex of enzyme IIAmannose and the histidine-containing phosphocarrier protein HPr from escherichia coli nmr, restrained regularized mean structure
1PFH 1995-11-14 THE PHOSPHORYLATED FORM OF THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR
3EZA 1999-05-25 COMPLEX OF THE AMINO TERMINAL DOMAIN OF ENZYME I AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
2LRK 2012-05-16 Solution Structures of the IIA(Chitobiose)-HPr complex of the N,N'-Diacetylchitobiose
3EZB 1999-12-16 COMPLEX OF THE AMINO TERMINAL DOMAIN OF ENZYME I AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI
1J6T 2002-11-13 COMPLEX OF ENZYME IIAMTL AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
3CCD 2008-10-21 1.0 1.0 A Structure of Post-Succinimide His15Asp HPr
1OPD 1997-08-20 1.5 HISTIDINE-CONTAINING PROTEIN (HPR), MUTANT WITH SER 46 REPLACED BY ASP (S46D)
1CM3 2000-05-17 1.6 HIS15ASP HPR FROM E. COLI
1CM2 2000-05-17 1.8 STRUCTURE OF HIS15ASP HPR AFTER HYDROLYSIS OF RINGED SPECIES.
1POH 1994-01-31 2.0 THE 2.0 ANGSTROMS RESOLUTION STRUCTURE OF ESCHERICHIA COLI HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR: A REDETERMINATION
4XWJ 2015-10-21 2.1 Histidine-containing phosphocarrier protein (HPr) and antisigma factor Rsd complex
2JEL 1998-05-27 2.5 JEL42 FAB/HPR COMPLEX
5YA1 2018-07-11 2.7 crystal structure of LsrK-HPr complex with ATP
5YA2 2018-07-11 2.7 Crystal structure of LsrK-HPr complex with ADP
5YA0 2018-07-11 3.0 Crystal structure of LsrK and HPr complex

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
98.8 Phosphocarrier protein HPr P16481 PTHP_KLEPN
100.0 Phosphocarrier protein HPr P0AA09 PTHP_SHIFL
100.0 Phosphocarrier protein HPr P0AA07 PTHP_SALTY
100.0 Phosphocarrier protein HPr P0AA08 PTHP_SALTI
100.0 Phosphocarrier protein HPr P0AA04 PTHP_ECOLI
100.0 Phosphocarrier protein HPr P0AA05 PTHP_ECOL6
100.0 Phosphocarrier protein HPr P0AA06 PTHP_ECO57