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

Study Summary

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

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 UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Peroxidase C2 P17179 PER2_ARMRU
100.0 Peroxidase C1A P00433 PER1A_ARMRU
93.1 Peroxidase C1A Q9SMU8 PER34_ARATH
91.2 Peroxidase C1A P24101 PER33_ARATH
91.2 Peroxidase C1A P15233 PER1C_ARMRU
90.8 Peroxidase C1A P15232 PER1B_ARMRU