Thermodynamic studies of the core histones: stability of the octamer subunits is not altered by removal of their terminal domains.


Abstract

We have investigated the role of the labile terminal domains of the core histones on the stability of the subunits of the protein core of the nucleosome by studying the thermodynamic behavior of the products of limited trypsin digestion of these subunits. The thermal stabilities of the truncated H2A-H2B dimer and the truncated (H3-H4)/(H3-H4)(2) system were studied by high-sensitivity differential scanning calorimetry and circular dichroism spectroscopy. The thermal denaturation of the truncated H2A-H2B dimer at pH 6.0 and low ionic strength is centered at 47.3 degrees C. The corresponding enthalpy change is 35 kcal/mol of 11.5 kDa monomer unit, and the heat capacity change upon unfolding is 1.2 kcal/(K mol of 11.5 kDa monomer unit). At pH 4.5 and low ionic strength, the truncated (H3-H4)/(H3-H4)(2) system, like its full-length counterpart, is quantitatively dissociated into two truncated H3-H4 dimers. The thermal denaturation of the truncated H3-H4 dimer is characterized by the presence of a single calorimetric peak centered at 60 degrees C. The enthalpy change is 25 kcal/mol of 10 kDa monomer unit, and the change in heat capacity upon unfolding is 0.5 kcal/(K mol of 10 kDa monomer unit). The thermal stabilities of both types of truncated dimers exhibit salt and pH dependencies similar to those of the full-length proteins. Finally, like their full-length counterparts, both truncated core histone dimers undergo thermal denaturation as highly cooperative units, without the involvement of any significant population of melting intermediates. Therefore, removal of the histone "tails" does not generally affect the thermodynamic behavior of the subunits of the core histone complex, indicating that the more centrally located regions of the histone fold and the extra-fold structured elements are primarily responsible for their stability and responses to parameters of their chemical microenvironment. Study holds ProTherm entries: 12238, 12239, 12240, 12241, 12242, 12243, 12244, 12245, 12246, 12247, 12248, 12249, 12250, 12251, 12252, 12253, 12254, 12255, 12256, 12257, 12258, 12259, 12260, 12261, 12262, 12263, 12264, 12265, 12266, 12267, 12268, 12269, 12270 Extra Details: Truncated H2A-H2B protein core; ionic strength; heat capacity change; chemical microenvironment

Submission Details

ID: Jnu9Ndop

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:43 p.m.

Version: 1

Publication Details
Karantza V;Freire E;Moudrianakis EN,Biochemistry (2001) Thermodynamic studies of the core histones: stability of the octamer subunits is not altered by removal of their terminal domains. PMID:11669650
Additional Information

Number of data points 33
Proteins Histone H2B 1/2/3/4/6 ; Histone H2A-IV
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: Tm prot_conc:142 micro M, buffers:glycylglycine: 10 mM, pH:4.0, ionic:NaCl: 5 mM ; Experimental Assay: Tm pH:4.0, buffers:glycylglycine: 10 mM, prot_conc:100 micro M, ionic:NaCl: 5 mM ; Experimental Assay: Tm buffers:glycylglycine: 10 mM, pH:4.0, prot_conc:60 micro M, ionic:NaCl: 5 mM ; Experimental Assay: Tm buffers:glycylglycine: 10 mM, pH:4.0, ionic:NaCl: 5 mM, prot_conc:30 micro M ; Experimental Assay: Tm prot_conc:15 micro M, buffers:glycylglycine: 10 mM, pH:4.0, ionic:NaCl: 5 mM ; Experimental Assay: Tm pH:5.0, ionic:NaCl: 50 mM, buffers:MES: 10 mM, prot_conc:32 micro M ; Experimental Assay: Tm pH:4.5, ionic:NaCl: 50 mM, buffers:MES: 10 mM, prot_conc:32 micro M ; Experimental Assay: Tm buffers:glycine: 10 mM, pH:4.0, ionic:NaCl: 50 mM, prot_conc:32 micro M ; Experimental Assay: Tm buffers:glycine: 10 mM, ionic:NaCl: 50 mM, pH:3.5, prot_conc:32 micro M ; Experimental Assay: Tm buffers:glycine: 10 mM, pH:3.0, ionic:NaCl: 50 mM, prot_conc:32 micro M ; Experimental Assay: Tm buffers:glycine: 10 mM, pH:2.5, ionic:NaCl: 50 mM, prot_conc:32 micro M ; Experimental Assay: Tm pH:4.5, ionic:NaCl: 50 mM, prot_conc:30 micro M, buffers:MES: 10 mM ; Experimental Assay: Tm ionic:NaCl: 25 mM, pH:4.5, prot_conc:30 micro M, buffers:MES: 10 mM ; Experimental Assay: Tm ionic:NaCl: 10 mM, pH:4.5, prot_conc:30 micro M, buffers:MES: 10 mM ; Experimental Assay: Tm pH:4.5, ionic:NaCl: 5 mM, prot_conc:30 micro M, buffers:MES: 10 mM ; Experimental Assay: Tm prot_conc:28 micro M, pH:4.2, ionic:NaCl: 12.5 mM, buffers:glycylglycine: 10 mM ; Experimental Assay: Tm prot_conc:28 micro M, ionic:NaCl: 100 mM, buffers:HEPES: 10 mM, pH:8.5 ; Experimental Assay: Tm prot_conc:28 micro M, ionic:NaCl: 100 mM, buffers:HEPES: 10 mM, pH:6.5 ; Experimental Assay: Tm pH:6.0, ionic:NaCl: 100 mM, prot_conc:28 micro M, buffers:HEPES: 10 mM ; Experimental Assay: Tm prot_conc:28 micro M, pH:5.5, ionic:NaCl: 100 mM, buffers:HEPES: 10 mM ; Experimental Assay: Tm prot_conc:28 micro M, pH:4.5, ionic:NaCl: 100 mM, buffers:HEPES: 10 mM ; Experimental Assay: Tm prot_conc:28 micro M, pH:4.0, ionic:NaCl: 100 mM, buffers:HEPES: 10 mM ; Experimental Assay: Tm prot_conc:28 micro M, ionic:NaCl: 100 mM, pH:3.5, buffers:HEPES: 10 mM ; Experimental Assay: Tm pH:7.5, ionic:NaCl: 1 M, prot_conc:28 micro M, buffers:HEPES: 10 mM ; Experimental Assay: Tm pH:7.5, prot_conc:28 micro M, buffers:HEPES: 10 mM, ionic:NaCl: 400 mM ; Experimental Assay: Tm pH:7.5, ionic:NaCl: 200 mM, prot_conc:28 micro M, buffers:HEPES: 10 mM ; Experimental Assay: Tm pH:7.5, ionic:NaCl: 100 mM, prot_conc:28 micro M, buffers:HEPES: 10 mM ; Experimental Assay: Tm pH:7.5, prot_conc:28 micro M, ionic:NaCl: 51 mM, buffers:HEPES: 10 mM ; Experimental Assay: Tm pH:7.5, prot_conc:28 micro M, ionic:NaCl: 21 mM, buffers:HEPES: 10 mM ; Experimental Assay: Tm pH:7.5, prot_conc:28 micro M, buffers:HEPES: 10 mM, ionic:NaCl: 11 mM ; Experimental Assay: Tm pH:7.5, ionic:NaCl: 1 mM, prot_conc:28 micro M, buffers:HEPES: 10 mM ; Experimental Assay: Tm ionic:NaCl: 25 mM, pH:6.0, prot_conc:28 micro M, buffers:imidazole: 10 mM
Libraries Mutations for sequence A:KSRSSRAGLQFPVGRVHRLLRKGNYAERVGAGAPVYLAAVLEYLTAEILELAGNAARDNKKTRIIPRHLQLAIRNDEELNKLLGKVTIAQGGVLP/B:SYSIYVYKVLKQVHPDTGISSKAMGSMNSFVNDIFERIAGLASRLAHYNKRSTITSREIQTAVRLLLPGELAKHAVSEGTKAVTKHTSSK/C:PGTVALREIRRYQKSTELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEASEAYLVGLFEDTNLCAIHAKRVTIMPKDIELARRIRGERA/D:QGITKPAIRRLARRGGVKRISGLIYEETRGVLKVFLENVIRDAVTYTEHAKRKTVTAMDVVYALKRQGRTLYGFGG
Sequence Assay Result Units