Thermodynamic stability of archaeal histones.


Abstract

The temperature, salt, and pH dependencies of unfolding of four recombinant (r) archaeal histones (rHFoB from the mesophile Methanobacterium formicicum, and rHMfA, rHMfB, and rHPyA1 from the hyperthermophiles Methanothermus fervidus and Pyrococcus strain GB-3a) have been determined by circular dichroism spectroscopy (CD) and differential scanning calorimetry (DSC). The thermal unfolding of these proteins is > 90% reversible, with concentration-dependent apparent Tm values and asymmetric unfolding transitions that are fit well by a two-state unfolding model in which a histone dimer unfolds to two random coil monomers. rHPyA1 dimers are stable in the absence of salt, whereas rHMfA, rHMfB, and rHFoB dimers unfold at 20 degrees C and pH 2 in solutions containing < 200 mM, < 400 mM, and < 1.5 M KCl, respectively. rHMfA, rHMfB, and rHFoB also experience significant cold denaturation in low salt concentrations and at low pH. The midpoint of thermal unfolding of a 1 M protein solution (T degree value) and the temperature dependency of the free energy of unfolding have been established for each histone, and both parameters correlate with the growth temperature of the originating archaeon. The changes in heat capacity upon unfolding are similar for the four histones, indicating that enhanced thermostability is not obtained by altering the curvature of the stability curve. Rather, the stability curves for the histones from the hyperthermophiles are displaced vertically to higher energies and laterally to higher Tmax values relative to the stability curve for rHFoB. The maximal free energies of unfolding for rHFoB, rHMfA, rHMfB, and rHPyA1 are 7.2, 15.5, 14.6, and 17.2 kcal/mol at 32, 35, 40, and 44 degrees C, respectively. T degree values for rHFoB, rHMfA, rHMfB, and rHPyA1 are 75, 104, 113, and 114 degrees C, respectively, at pH 5 in 0.2 M KCl. Structural features within the conserved histone fold that might confer these stability differences are discussed. Study holds ProTherm entries: 8740, 8741, 8742, 8743, 8744, 8745, 8746, 8747, 8748, 8749, 8750, 8751, 8752, 8753, 8754, 8755, 8756, 8757, 8758, 8759, 8760, 8761, 8762, 8763, 8764, 8765, 8766, 8767, 8768, 8769, 8770, 8771, 8772, 8773, 8774, 8775, 8776 Extra Details: rHMfA two-state unfolding model; histone dimer; temperature dependency;,conserved histone fold

Submission Details

ID: kvvfAagh

Submitter: Connie Wang

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

Version: 1

Publication Details
Li WT;Grayling RA;Sandman K;Edmondson S;Shriver JW;Reeve JN,Biochemistry (1998) Thermodynamic stability of archaeal histones. PMID:9692945
Additional Information

Study Summary

Number of data points 61
Proteins Archaeal histone A1 ; DNA-binding protein HMf-2 ; DNA-binding protein HMf-1 ; Archaeal histone B
Unique complexes 3
Assays/Quantities/Protocols Experimental Assay: Tm pH:7.5, buffers:phosphate: 25 mM, prot_conc:1.8-5 micro M ; Experimental Assay: Tm prot_conc:1.8-5 micro M, buffers:glycine: 25 mM, pH:3.7 ; Experimental Assay: Tm prot_conc:1.8-5 micro M, pH:3.3, buffers:glycine: 25 mM ; Experimental Assay: Tm prot_conc:1.8-5 micro M, buffers:glycine: 25 mM, pH:2.7 ; Experimental Assay: Tm prot_conc:1.8-5 micro M, pH:2.5, buffers:glycine: 25 mM ; Experimental Assay: Tm pH:6.0, prot_conc:1.8-5 micro M, ionic:KCl: 1.0 M, buffers:MES: 25 mM ; Experimental Assay: Tm prot_conc:1.8-5 micro M, pH:5.0, ionic:KCl: 1.0 M, buffers:acetate: 25 mM ; Experimental Assay: Tm prot_conc:1.8-5 micro M, pH:4.0, ionic:KCl: 1.0 M, buffers:glycine: 25 mM ; Experimental Assay: Tm prot_conc:1.8-5 micro M, pH:3.0, ionic:KCl: 1.0 M, buffers:glycine: 25 mM ; Experimental Assay: Tm prot_conc:1.8-5 micro M, ionic:KCl: 1.0 M, pH:2.0, buffers:glycine: 25 mM ; Experimental Assay: Tm pH:6.0, prot_conc:1.8-5 micro M, buffers:MES: 25 mM ; Experimental Assay: Tm prot_conc:1.8-5 micro M, pH:5.0, buffers:acetate: 25 mM ; Experimental Assay: Tm prot_conc:1.8-5 micro M, pH:4.0, buffers:glycine: 25 mM ; Experimental Assay: Tm prot_conc:1.8-5 micro M, pH:3.0, buffers:glycine: 25 mM ; Experimental Assay: dHcal pH:6.0, ionic:KCl: 1.0 M, buffers:MES: 25 mM ; Experimental Assay: Tm pH:6.0, ionic:KCl: 1.0 M, buffers:MES: 25 mM, prot_conc:150-500 micro M ; Experimental Assay: dHvH pH:6.0, ionic:KCl: 1.0 M, buffers:MES: 25 mM ; Experimental Assay: dHcal pH:5.0, ionic:KCl: 1.0 M, buffers:acetate: 25 mM ; Experimental Assay: Tm pH:5.0, ionic:KCl: 1.0 M, prot_conc:150-500 micro M, buffers:acetate: 25 mM ; Experimental Assay: dHvH pH:5.0, ionic:KCl: 1.0 M, buffers:acetate: 25 mM ; Experimental Assay: dHcal pH:4.0, ionic:KCl: 1.0 M, buffers:glycine: 25 mM ; Experimental Assay: Tm ionic:KCl: 1.0 M, pH:4.0, buffers:glycine: 25 mM, prot_conc:150-500 micro M ; Experimental Assay: dHvH pH:4.0, ionic:KCl: 1.0 M, buffers:glycine: 25 mM ; Experimental Assay: dHcal pH:3.0, ionic:KCl: 1.0 M, buffers:glycine: 25 mM ; Experimental Assay: Tm ionic:KCl: 1.0 M, pH:3.0, buffers:glycine: 25 mM, prot_conc:150-500 micro M ; Experimental Assay: dHvH pH:3.0, ionic:KCl: 1.0 M, buffers:glycine: 25 mM ; Experimental Assay: dHcal ionic:KCl: 1.0 M, pH:2.0, buffers:glycine: 25 mM ; Experimental Assay: Tm pH:2.0, ionic:KCl: 1.0 M, buffers:glycine: 25 mM, prot_conc:150-500 micro M ; Experimental Assay: dHvH ionic:KCl: 1.0 M, pH:2.0, buffers:glycine: 25 mM ; Experimental Assay: dHcal pH:6.0, buffers:MES: 25 mM, ionic:KCl: 0.2 M ; Experimental Assay: Tm pH:6.0, buffers:MES: 25 mM, prot_conc:150-500 micro M ; Experimental Assay: dHvH pH:6.0, buffers:MES: 25 mM, ionic:KCl: 0.2 M ; Experimental Assay: dHcal pH:5.0, ionic:KCl: 0.2 M, buffers:acetate: 25 mM ; Experimental Assay: Tm pH:5.0, prot_conc:150-500 micro M, buffers:acetate: 25 mM ; Experimental Assay: dHvH pH:5.0, ionic:KCl: 0.2 M, buffers:acetate: 25 mM ; Experimental Assay: dHcal pH:4.0, buffers:glycine: 25 mM, ionic:KCl: 0.2 M ; Experimental Assay: Tm pH:4.0, buffers:glycine: 25 mM, prot_conc:150-500 micro M ; Experimental Assay: dHvH pH:4.0, buffers:glycine: 25 mM, ionic:KCl: 0.2 M ; Experimental Assay: dHcal pH:3.0, buffers:glycine: 25 mM, ionic:KCl: 0.2 M ; Experimental Assay: Tm pH:3.0, buffers:glycine: 25 mM, prot_conc:150-500 micro M ; Experimental Assay: dHvH pH:3.0, buffers:glycine: 25 mM, ionic:KCl: 0.2 M
Libraries Mutations for sequence MELPIAPIGRIIKDAGAERVSDDARITLAKILEEMGRDIASEAIKLARHAGRKTIKAEDIELAVRRFKK ; Mutations for sequence MAELPIAPVGRIIKNAGAPRVSDDARDALAKVLEEMGEGIAAEAVKLAKHAGRKTVKASDIEMAVKAA ; Mutations for sequence MGELPIAPVDRLIRKAGAQRVSEQAAKVLAEHLEEKAIEIAKKAVDLAKHAGRKTVKVEDIKLAIKS

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
1BFM 1996-01-29 HISTONE B FROM METHANOTHERMUS FERVIDUS
1B67 2000-01-17 1.48 CRYSTAL STRUCTURE OF THE HISTONE HMFA FROM METHANOTHERMUS FERVIDUS
1HTA 1999-03-23 1.55 CRYSTAL STRUCTURE OF THE HISTONE HMFA FROM METHANOTHERMUS FERVIDUS
1A7W 1999-03-23 1.55 CRYSTAL STRUCTURE OF THE HISTONE HMFB FROM METHANOTHERMUS FERVIDUS
1B6W 2000-01-17 2.05 CRYSTAL STRUCTURE OF THE SELENOMETHIONINE VARIANT OF HISTONE HMFB FROM METHANOTHERMUS FERVIDUS
1KU5 2003-08-26 2.3 Crystal Structure of recombinant histone HPhA from hyperthermophilic archaeon Pyrococcus horikoshii OT3
5T5K 2017-08-23 4.0 Structure of histone-based chromatin in Archaea

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Archaeal histone A1 P48782 HFO1_METFO
100.0 DNA-binding protein HMf-2 P19267 HMFB_METFE
100.0 DNA-binding protein HMf-1 P48781 HMFA_METFE
91.0 Archaeal histone B O74098 HARA_PYRHO
91.0 Archaeal histone B P50486 HARB_PYRSG
91.0 Archaeal histone B O74092 HARB_PYRHO
92.5 Archaeal histone B Q9V1F5 HARB_PYRAB
100.0 Archaeal histone B O59627 HARB_PYRFU