Conformational stability of HPr: the histidine-containing phosphocarrier protein from Bacillus subtilis.


Abstract

The conformational stability of the histidine-containing phosphocarrier protein (HPr) from Bacillus subtilis has been determined using a combination of thermal unfolding and solvent denaturation experiments. The urea-induced denaturation of HPr was monitored spectroscopically at fixed temperatures and thermal unfolding was performed in the presence of fixed concentrations of urea. These data were analyzed in several different ways to afford a measure of the cardinal parameters (delta Hg, Tg, delta Sg, and delta Cp) that describe the thermodynamics of folding for HPr. The method of Pace and Laurents (Pace CN, Laurents DV, 1989, Biochemistry 28:2520-2525) was used to estimate delta Cp as was a global analysis of the thermal- and urea-induced unfolding data. Each method used to analyze the data gives a similar value for delta Cp (1,170 +/- 50 cal mol-1K-1). Despite the high melting temperature for HPr (Tg = 73.5 degrees C), the maximum stability of the protein, which occurs at 26 degrees C, is quite modest (delta Gs = 4.2 kcal mol-1). In the presence of moderate concentrations of urea, HPr exhibits cold denaturation, and thus a complete stability curve for HPr, including a measure of delta Cp, can be achieved using the method of Chen and Schellman (Chen B, Schellman JA, 1989, Biochemistry 28:685-691). A comparison of the different methods for the analysis of solvent denaturation curves is provided and the effects of urea on the thermal stability of this small globular protein are discussed. The methods presented will be of general utility in the characterization of the stability curve for many small proteins. Study holds ProTherm entries: 7284, 7285, 7286, 7287, 7288, 7289, 7290, 7291, 7292 Extra Details: cold denaturation; conformational stability; protein folding

Submission Details

ID: k2kPady

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:33 p.m.

Version: 1

Publication Details
Scholtz JM,Protein Sci. (1995) Conformational stability of HPr: the histidine-containing phosphocarrier protein from Bacillus subtilis. PMID:7773175
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
2HID 1996-11-08 REFINED NMR STRUCTURE OF PHOSPHOCARRIER HISTIDINE CONTAINING PROTEIN FROM BACILLUS SUBTILIS
1JEM 1997-07-23 NMR STRUCTURE OF HISTIDINE PHOSPHORYLATED FORM OF THE PHOSPHOCARRIER HISTIDINE CONTAINING PROTEIN FROM BACILLUS SUBTILIS, NMR, 25 STRUCTURES
2HPR 1993-01-15 2.0 HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR MUTANT WITH MET 51 REPLACED BY VAL AND SER 83 REPLACED BY CYS (M51V, S83C)
1SPH 1995-02-07 2.0 REFINED STRUCTURES OF THE ACTIVE S83C AND IMPAIRED S46D HPRS: EVIDENCE THAT PHOSPHORYLATION DOES NOT REQUIRE A BACKBONE CONFORMATIONAL TRANSITION
2FEP 2006-06-27 2.45 Structure of truncated CcpA in complex with P-Ser-HPr and Sulfate ions
1KKM 2002-08-28 2.8 L.casei HprK/P in complex with B.subtilis P-Ser-HPr
1KKL 2002-08-28 2.8 L.casei HprK/P in complex with B.subtilis HPr
3OQM 2010-12-08 2.96 structure of ccpa-hpr-ser46p-ackA2 complex
3OQO 2011-10-26 2.97 Ccpa-hpr-ser46p-syn cre
3OQN 2010-12-08 3.3 Structure of ccpa-hpr-ser46-p-gntr-down cre

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Phosphocarrier protein HPr P08877 PTHP_BACSU