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 PDB Entries

Structure ID Release Date Resolution Structure Title
1GWU 2003-03-28 1.31 RECOMBINANT HORSERADISH PEROXIDASE C1A ALA140GLY
7ATJ 2000-01-14 1.47 RECOMBINANT HORSERADISH PEROXIDASE C1A COMPLEX WITH CYANIDE AND FERULIC ACID
1W4W 2005-01-19 1.55 Ferric horseradish peroxidase C1A in complex with formate
1HCH 2002-07-19 1.57 Structure of horseradish peroxidase C1A compound I
1H5E 2002-05-27 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (11-22% dose)
1H5D 2002-05-27 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (0-11% dose)
1H5L 2002-06-21 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (89-100% dose)
1H5M 2002-05-27 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (0-100% dose)
1H5I 2002-05-27 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (56-67% dose)
1H5J 2002-05-27 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (67-78% dose)
1H5A 2002-06-18 1.6 STRUCTURE OF FERRIC HORSERADISH PEROXIDASE C1A IN COMPLEX WITH ACETATE
1H57 2002-06-17 1.6 Structure of horseradish peroxidase C1A compound III
1H5G 2002-05-27 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (33-44% dose)
1H5F 2002-05-27 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (22-33% dose)
1H5K 2002-05-27 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (78-89% dose)
1W4Y 2005-01-19 1.6 Ferrous horseradish peroxidase C1A in complex with carbon monoxide
1H5H 2002-06-21 1.6 X-ray induced reduction of horseradish peroxidase C1A Compound III (44-56% dose)
1H55 2002-06-18 1.61 STRUCTURE OF HORSERADISH PEROXIDASE C1A COMPOUND II
1H5C 2002-06-18 1.62 X-ray induced reduction of horseradish peroxidase C1A Compound III (100-200% dose)
2YLJ 2012-06-13 1.69 Horse Radish Peroxidase, mutant S167Y
1GWT 2003-03-28 1.7 RECOMBINANT HORSERADISH PEROXIDASE C1A PHE221MET
1H58 2002-06-18 1.7 STRUCTURE OF FERROUS HORSERADISH PEROXIDASE C1A
2ATJ 1998-01-28 2.0 RECOMBINANT HORSERADISH PEROXIDASE COMPLEX WITH BENZHYDROXAMIC ACID
1KZM 2002-10-09 2.0 Distal Heme Pocket Mutant (R38S/H42E) of Recombinant Horseradish Peroxidase C (HRP C).
6ATJ 2000-01-14 2.0 RECOMBINANT HORSERADISH PEROXIDASE C COMPLEX WITH FERULIC ACID
1GWO 2003-03-28 2.07 Recombinant horseradish peroxidase C1A ALA170GLN
1GW2 2003-03-28 2.15 RECOMBINANT HORSERADISH PEROXIDASE C1A THR171SER IN COMPLEX WITH FERULIC ACID
1ATJ 1998-02-04 2.15 RECOMBINANT HORSERADISH PEROXIDASE C1A
3ATJ 1999-04-28 2.2 HEME LIGAND MUTANT OF RECOMBINANT HORSERADISH PEROXIDASE IN COMPLEX WITH BENZHYDROXAMIC ACID
1GX2 2003-03-28 2.2 Recombinant horseradish peroxidase Phe209Ser complex with benzhydroxamic acid
4ATJ 2002-10-02 2.5 DISTAL HEME POCKET MUTANT (H42E) OF RECOMBINANT HORSERADISH PEROXIDASE IN COMPLEX WITH BENZHYDROXAMIC ACID

Relevant UniProtKB Entries

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