Denaturation of horseradish peroxidase with urea and guanidine hydrochloride.


Abstract

Favourable effects of urea and guanidine hydrochloride (Gdn HCl) on solubilization of the polar, non-polar and peptide groups of horseradish peroxidase (HRP), an example of a globular protein, provide the driving force for unfolding of HRP, in a reversible two-state process. The intrinsic or conformational stability of HRP at various pH values and temperatures has been estimated by the linear extrapolation method (LEM), a denaturant binding model (DBM) and Tanford's model. There is good agreement between these methods. Tanford's model shows that urea interacts with non-polar groups to a greater extent than Gdn HCl does, whereas Gdn HCl interacts more effectively with the peptide groups of HRP. Study holds ProTherm entries: 12316, 12317, 12318, 12319, 12320, 12321, 12322, 12323, 12324, 12325, 12326, 12327, 12328, 12329, 12330, 12331, 12332, 12333, 12334, 12335, 12336, 12337, 12338, 12339 Extra Details: horseradish peroxidase; denaturation; urea; guanidine hydrochloride

Submission Details

ID: rPwf5my7

Submitter: Connie Wang

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

Version: 1

Publication Details
Moosavi-Movahedi AA;Nazari K,Int. J. Biol. Macromol. (1995) Denaturation of horseradish peroxidase with urea and guanidine hydrochloride. PMID:7772563
Additional Information

Number of data points 48
Proteins Peroxidase C2 ; Peroxidase C1A
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: m temp:30.0 C, pH:10.0 ; Experimental Assay: dG_H2O temp:30.0 C, pH:10.0 ; Experimental Assay: m temp:30.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: dG_H2O temp:30.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: m temp:30.0 C, pH:3.2 ; Experimental Assay: dG_H2O temp:30.0 C, pH:3.2 ; Experimental Assay: m temp:27.0 C, pH:10.0 ; Experimental Assay: dG_H2O temp:27.0 C, pH:10.0 ; Experimental Assay: m temp:27.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: dG_H2O temp:27.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: m temp:27.0 C, pH:3.2 ; Experimental Assay: dG_H2O temp:27.0 C, pH:3.2 ; Experimental Assay: m temp:25.0 C, pH:10.0 ; Experimental Assay: dG_H2O temp:25.0 C, pH:10.0 ; Experimental Assay: m pH:6.4, buffers:sodium phosphate: 2.5 mM, temp:25.0 C ; Experimental Assay: dG_H2O pH:6.4, temp:25.0 C, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: m pH:3.2, temp:25.0 C ; Experimental Assay: dG_H2O temp:25.0 C, pH:3.2 ; Experimental Assay: m temp:22.0 C, pH:10.0 ; Experimental Assay: dG_H2O temp:22.0 C, pH:10.0 ; Experimental Assay: m temp:22.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: dG_H2O temp:22.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: m temp:22.0 C, pH:3.2 ; Experimental Assay: dG_H2O temp:22.0 C, pH:3.2 ; Experimental Assay: m temp:30.0 C, pH:10.0 ; Experimental Assay: dG_H2O temp:30.0 C, pH:10.0 ; Experimental Assay: m temp:30.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: dG_H2O temp:30.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: m temp:30.0 C, pH:3.2 ; Experimental Assay: dG_H2O temp:30.0 C, pH:3.2 ; Experimental Assay: m temp:27.0 C, pH:10.0 ; Experimental Assay: dG_H2O temp:27.0 C, pH:10.0 ; Experimental Assay: m temp:27.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: dG_H2O temp:27.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: m temp:27.0 C, pH:3.2 ; Experimental Assay: dG_H2O temp:27.0 C, pH:3.2 ; Experimental Assay: m temp:25.0 C, pH:10.0 ; Experimental Assay: dG_H2O temp:25.0 C, pH:10.0 ; Experimental Assay: m pH:6.4, buffers:sodium phosphate: 2.5 mM, temp:25.0 C ; Experimental Assay: dG_H2O pH:6.4, temp:25.0 C, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: m pH:3.2, temp:25.0 C ; Experimental Assay: dG_H2O temp:25.0 C, pH:3.2 ; Experimental Assay: m temp:22.0 C, pH:10.0 ; Experimental Assay: dG_H2O temp:22.0 C, pH:10.0 ; Experimental Assay: m temp:22.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: dG_H2O temp:22.0 C, pH:6.4, buffers:sodium phosphate: 2.5 mM ; Experimental Assay: m temp:22.0 C, pH:3.2 ; Experimental Assay: dG_H2O temp:22.0 C, pH:3.2
Libraries Mutations for sequence QLTPTFYDNSCPNVSNIVRDTIVNELRSDPRIAASILRLHFHDCFVNGCDASILLDNTTSFRTEKDAFGNANSARGFPVIDRMKAAVESACPRTVSCADLLTIAAQQSVTLAGGPSWRVPLGRRDSLQAFLDLANANLPAPFFTLPQLKDSFRNVGLNRSSDLVALSGGHTFGKNQCRFIMDRLYNFSNTGLPDPTLNTTYLQTLRGLCPLNGNLSALVDFDLRTPTIFDNKYYVNLEEQKGLIQSDQELFSSPNATDTIPLVRSFANSTQTFFNAFVEAMDRMGNITPLTGTQGQIRLNCRVVNS
Sequence Assay Result Units