Guanidine hydrochloride-induced folding of proteins.


Abstract

Guanidine hydrochloride (Gdn-HCl) is the most commonly used denaturant for proteins. Contrary to expectation, we found that Gdn-HCl at low concentrations refolds acid-unfolded apomyoglobin and cytochrome c, stabilizing the molten globule state, i.e. a compact denatured state with a significant amount of secondary structure, but substantially disordered tertiary structure. A further increase in Gdn-HCl concentration, above 1 M, caused co-operative unfolding of the molten globule state. Similar sequential folding and unfolding transitions were also observed at neutral pH with a synthetic amphiphilic peptide consisting of Lys and Leu residues, indicating the generality of the phenomenon. Although the Gdn-HCl-induced refolding and unfolding transitions were puzzling at first glance, we show that they are readily interpreted in terms of the differential action of Gdn-HCl. We also show that the comparison of the unfolding curves for the molten globule and native states provides a measure of the buried surface area upon formation of the molten globule state. Study holds ProTherm entries: 9244, 9245 Extra Details: apomyoglobin; cytochrome c; guanidine hydrochloride;,molten globule; protein folding

Submission Details

ID: VRhgqxv9

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:37 p.m.

Version: 1

Publication Details
Hagihara Y;Aimoto S;Fink AL;Goto Y,J. Mol. Biol. (1993) Guanidine hydrochloride-induced folding of proteins. PMID:8389881
Additional Information

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
1M60 2002-08-07 Solution Structure of Zinc-substituted cytochrome c
1GIW 1998-12-09 SOLUTION STRUCTURE OF REDUCED HORSE HEART CYTOCHROME C, NMR, MINIMIZED AVERAGE STRUCTURE
2N3B 2015-10-28 Structure of oxidized horse heart cytochrome c encapsulated in reverse micelles
1FI7 2000-08-23 Solution structure of the imidazole complex of cytochrome C
1LC2 2003-06-03 Solution Structure Of Reduced Horse Heart Cytochrome c in 30% Acetonitrile Solution, NMR 30 Structures
1OCD 1997-06-16 CYTOCHROME C (OXIDIZED) FROM EQUUS CABALLUS, NMR, MINIMIZED AVERAGE STRUCTURE
1LC1 2003-06-03 Solution Structure Of Reduced Horse Heart Cytochrome c in 30% Acetonitrile Solution, NMR Minimized Average Structure
2GIW 1998-12-09 SOLUTION STRUCTURE OF REDUCED HORSE HEART CYTOCHROME C, NMR, 40 STRUCTURES
2FRC 1997-07-29 CYTOCHROME C (REDUCED) FROM EQUUS CABALLUS, NMR, MINIMIZED AVERAGE STRUCTURE
1FI9 2000-08-23 SOLUTION STRUCTURE OF THE IMIDAZOLE COMPLEX OF CYTOCHROME C
1AKK 1997-09-17 SOLUTION STRUCTURE OF OXIDIZED HORSE HEART CYTOCHROME C, NMR, MINIMIZED AVERAGE STRUCTURE
5ZKV 2019-05-22 Solution structure of molten globule state of L94G mutant of horse cytochrome-c
1I5T 2001-03-21 SOLUTION STRUCTURE OF CYANOFERRICYTOCHROME C
3VM9 2012-09-12 1.05 Dimeric horse myoglobin
4TWV 2016-05-04 1.06 Horse heart myoglobin mutant (K45E/K63E/K96E) with Zn-deuteroporphyrin IX
4TWU 2016-05-04 1.08 Horse heart myoglobin mutant (D44K/D60K/E85K) with Zn-deuteroporphyrin IX
5C0Z 2016-09-21 1.12 The structure of oxidized rat cytochrome c at 1.13 angstroms resolution
4NS2 2014-12-24 1.18 Crystal structure of Horse heart ferric myoglobin; D44K/D60K/E85K mutant
2V1F 2007-06-12 1.2 Crystal structure of radiation-induced myoglobin compound II - intermediate H at pH 8.7
2FRF 2006-05-30 1.2 Horse Heart Myoglobin, Nitrite Adduct, Crystal Soak
5AZR 2016-01-20 1.2 Crystal structure of aqua-cobalt(III) tetradehydrocorrin in the heme pocket of horse heart myoglobin
2V1I 2007-06-12 1.2 Crystal structure of radiation-induced metmyoglobin - aqua ferrous myoglobin at pH 6.8
3RJ6 2012-05-09 1.23 Crystal Structure of Horse heart ferric myoglobin; K45E/K63E/K96E mutant
2V1K 2007-06-12 1.25 Crystal structure of ferrous deoxymyoglobin at pH 6.8
2FRJ 2006-05-30 1.3 Nitrosyl Horse Heart Myoglobin, Nitrite/Dithionite Method
2V1E 2007-06-12 1.3 Crystal structure of radiation-induced myoglobin compound II - intermediate H at pH 6.8
5DF5 2016-09-14 1.3 The structure of oxidized rat cytochrome c (T28E) at 1.30 angstroms resolution.
2V1H 2007-06-12 1.3 Crystal structure of radiation-induced metmyoglobin - aqua ferrous myoglobin at pH 5.2
2VLX 2009-02-10 1.3 Crystal structure of peroxymyoglobin generated by cryoradiolytic reduction of myoglobin compound III
3WFT 2014-12-03 1.3 Crystal structure of horse heart myoglobin reconstituted with cobalt(II) tetradehydrocorrin
2FRK 2006-05-30 1.3 Nitrosyl Horse Heart Myoglobin, Nitric Oxide Gas Method
2V1G 2007-06-12 1.35 Crystal structure of radiation-induced myoglobin compound II - intermediate H at pH 5.2
1GJN 2002-03-01 1.35 Hydrogen Peroxide Derived Myoglobin Compound II at pH 5.2
3WFU 2014-12-03 1.35 Crystal structure of horse heart myoglobin reconstituted with cobalt(I) tetradehydrocorrin
5C9M 2016-09-21 1.36 The structure of oxidized rat cytochrome c (T28A) at 1.362 angstroms resolution.
5AZQ 2016-01-20 1.4 Crystal structure of cyano-cobalt(III) tetradehydrocorrin in the heme pocket of horse heart myoglobin
1DWT 2000-03-03 1.4 Photorelaxed horse heart MYOGLOBIN CO complex
2V1J 2007-06-12 1.4 Crystal structure of radiation-induced metmyoglobin - aqua ferrous myoglobin at pH 8.7
5ZZE 2019-02-20 1.42 Crystal structure of horse myoglobin crystallized by ammonium sulfate
1DWS 2000-03-03 1.45 PHOTOLYZED CARBONMONOXY MYOGLOBIN (HORSE HEART)
1DWR 2000-03-03 1.45 MYOGLOBIN (HORSE HEART) WILD-TYPE COMPLEXED WITH CO
2B4Z 2005-10-11 1.5 Crystal structure of cytochrome C from bovine heart at 1.5 A resolution.
4DC7 2013-01-23 1.5 Crystal Structure of Myoglobin Exposed to Excessive SONICC Imaging Laser Dose.
4DC8 2013-01-23 1.5 Crystal Structure of Myoglobin Unexposed to Excessive SONICC Imaging Laser Dose.
2VLZ 2008-01-29 1.5 Crystal structure of peroxymyoglobin generated by cryoradiolytic reduction of myoglobin compound III
5YL3 2017-12-27 1.5 Crystal structure of horse heart myoglobin reconstituted with manganese porphycene in resting state at pH 8.5
3LR9 2010-07-07 1.55 X-ray photogenerated ferrous horse heart myoglobin, nitrite adduct
5YCJ 2018-09-19 1.58 Ancestral myoglobin aMbWb' of Basilosaurus relative (polyphyly) imidazole-ligand
5D5R 2015-09-16 1.6 Horse-heart myoglobin - deoxy state
1NZ3 2003-04-08 1.6 K45E-K63E Variant of Horse Heart Myoglobin
2O5S 2007-10-16 1.6 Cobalt horse heart myoglobin, nitrite modified
2O5O 2007-10-16 1.6 Manganese horse heart myoglobin, nitrite modified
2O5T 2007-10-16 1.6 Cobalt horse heart myoglobin, oxidized
2VLY 2008-01-29 1.6 Crystal structure of myoglobin compound III (radiation-induced)
2FRI 2006-05-30 1.6 Horse Heart Myoglobin, Nitrite Adduct, Co-crystallized
3LR7 2010-07-07 1.6 Ferric horse heart myoglobin, nitrite adduct
2VM0 2008-01-29 1.6 Crystal structure of radiation-induced myoglobin compound II generated after annealing of peroxymyoglobin
2O58 2007-10-16 1.65 Horse heart met manganese myoglobin
2O5M 2007-10-16 1.65 Manganese horse heart myoglobin, azide modified
3V2V 2012-11-21 1.65 Nitrite Bound Chlorin Substituted Myoglobin- Method 1
3V2Z 2012-11-21 1.65 Nitrite Bound Chlorin Substituted Myoglobin- Method 2
1NPG 2003-11-11 1.7 MYOGLOBIN (HORSE HEART) WILD-TYPE COMPLEXED WITH NITROSOETHANE
1MWC 1998-08-19 1.7 WILD TYPE MYOGLOBIN WITH CO
1XCH 1997-09-17 1.7 MYOGLOBIN (HORSE HEART) MUTANT WITH LEU 104 REPLACED BY ASN (L104N)
2O5L 2007-10-16 1.7 Manganese horse heart myoglobin, methanol modified
3VAU 2012-04-11 1.7 Myoglobin nitrite structure: nitriheme modified
1WLA 1998-01-14 1.7 MYOGLOBIN (HORSE HEART) RECOMBINANT WILD-TYPE
1RSE 1996-12-23 1.7 MYOGLOBIN (HORSE HEART) MUTANT WITH SER 92 REPLACED BY ASP (S92D)
1NZ5 2003-04-08 1.7 The Horse heart myoglobin variant K45E/K63E complexed with Manganese
1HRM 1995-01-26 1.7 THE PROXIMAL LIGAND VARIANT HIS93TYR OF HORSE HEART MYOGLOBIN
6FF5 2018-03-21 1.74 X-ray structure of bovine heart cytochrome c at high ionic strength
1MYG 1994-01-31 1.75 HIGH RESOLUTION X-RAY STRUCTURES OF PIG METMYOGLOBIN AND TWO CD3 MUTANTS MB(LYS45-> ARG) AND MB(LYS45-> SER)
3O1Y 2012-01-25 1.75 Electron transfer complexes: Experimental mapping of the redox-dependent cytochrome c electrostatic surface
5CMV 2015-09-16 1.8 Ultrafast dynamics in myoglobin: dark-state, CO-ligated structure
5CNF 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 50 ps time delay
5Z7E 2019-01-30 1.8 Horse Heart Myoglobin Mutant - H93M
5CNG 2015-09-16 1.8 ultrafast dynamics in myoglobin: 150 ps time delay
5CNE 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 10 ps time delay
5CN5 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0 ps time delay
3WUI 2014-07-16 1.8 Dimeric horse cytochrome c formed by refolding from molten globule state
5CN4 2015-09-16 1.8 Ultrafast dynamics in myoglobin: -0.1 ps time delay
1BJE 1998-01-28 1.8 H64T VARIANT OF MYOGLOBIN (HORSE HEART) RECOMBINANT WILD-TYPE COMPLEXED WITH AZIDE
1MWD 1998-08-19 1.8 WILD TYPE DEOXY MYOGLOBIN
5CNC 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.6 ps time delay
5CN6 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.1 ps time delay
5CN9 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.4 ps time delay
1NZ4 2003-04-08 1.8 The horse heart myoglobin variant K45E/K63E complexed with Cadmium
5CNB 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.5 ps time delay
5CND 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 3 ps time delay
1WEJ 1998-12-09 1.8 IGG1 FAB FRAGMENT (OF E8 ANTIBODY) COMPLEXED WITH HORSE CYTOCHROME C AT 1.8 A RESOLUTION
5CN8 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.3 ps time delay
5CN7 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.2 ps time delay
1M6M 1998-08-19 1.8 V68N MET MYOGLOBIN
3BA2 2008-11-11 1.8 Cyanide bound Chlorin substituted Myoglobin
3WC8 2013-12-11 1.8 Dimeric horse cytochrome c obtained by refolding with desalting method
1HSY 1995-02-27 1.9 ORIGIN OF THE PH-DEPENDENT SPECTROSCOPIC PROPERTIES OF PENTACOORDINATE METMYOGLOBIN VARIANTS
1MYH 1994-01-31 1.9 HIGH RESOLUTION X-RAY STRUCTURES OF PIG METMYOGLOBIN AND TWO CD3 MUTANTS MB(LYS45-> ARG) AND MB(LYS45-> SER)
1YMB 1994-01-31 1.9 HIGH RESOLUTION STUDY OF THE THREE-DIMENSIONAL STRUCTURE OF HORSE HEART METMYOGLOBIN
5Z7F 2019-01-30 1.9 Horse Heart Myoglobin Mutant -V68A/I107Y-Sulfide Derivative
1M6C 1998-08-19 1.9 V68N MYOGLOBIN WITH CO
1NPF 2003-11-11 1.9 MYOGLOBIN (HORSE HEART) WILD-TYPE COMPLEXED WITH NITRIC OXIDE
1MYJ 1994-01-31 1.9 DISTAL POLARITY IN LIGAND BINDING TO MYOGLOBIN: STRUCTURAL AND FUNCTIONAL CHARACTERIZATION OF A THREONINE68(E11) MUTANT
3HEN 2009-12-29 1.9 Ferric Horse Heart Myoglobin; H64V/V67R Mutant
2O5Q 2007-10-16 1.9 Manganese horse heart myoglobin, nitric oxide modified
3O20 2012-01-25 1.9 Electron transfer complexes:experimental mapping of the Redox-dependent Cytochrome C electrostatic surface
2NSR 2007-11-06 1.9 Nitromethane Modified Horse Heart Myoglobin
3RJN 2012-05-09 1.9 Horse heart myoglobin: D44K/D60K mutant with zinc (II) -deuteroporphyrin dimethyl ester
1HRC 1994-11-01 1.9 HIGH-RESOLUTION THREE-DIMENSIONAL STRUCTURE OF HORSE HEART CYTOCHROME C
1NZ2 2003-04-08 1.9 K45E Variant of Horse Heart Myoglobin
1MNO 1998-08-19 1.95 V68N MYOGLOBIN OXY FORM
3HEP 2009-12-29 1.95 Ferric Horse Heart Myoglobin; H64V Mutant, Nitrite Modified
5YCI 2018-09-19 1.97 Ancestral myoglobin aMbWb' of Basilosaurus relative (polyphyly)
1MDN 1998-09-30 1.98 WILD TYPE MYOGLOBIN WITH CO
2YBB 2011-10-19 19.0 Fitted model for bovine mitochondrial supercomplex I1III2IV1 by single particle cryo-EM (EMD-1876)
2O5B 2007-10-16 2.0 Manganese horse heart myoglobin, reduced
1YMA 1994-01-31 2.0 STRUCTURAL CHARACTERIZATION OF HEME LIGATION IN THE HIS64-->TYR VARIANT OF MYOGLOBIN
3WYO 2014-11-19 2.0 Heterodimeric myoglobin formed by domain swapping
2NSS 2007-11-06 2.0 Nitrobenzene Modified Horse Heart Myoglobin
5IY5 2017-01-11 2.0 Electron transfer complex of cytochrome c and cytochrome c oxidase at 2.0 angstrom resolution
1YMC 1994-01-31 2.0 THREE-DIMENSIONAL STRUCTURE OF CYANOMET-SULFMYOGLOBIN C
1MYI 1994-01-31 2.0 HIGH RESOLUTION X-RAY STRUCTURES OF PIG METMYOGLOBIN AND TWO CD3 MUTANTS MB(LYS45-> ARG) AND MB(LYS45-> SER)
3HEO 2009-12-29 2.0 Ferric Horse Heart Myoglobin; H64V/V67R mutant, Nitrite Modified
3HC9 2009-12-29 2.0 Ferric Horse Heart Myoglobin; H64V mutant
1AZI 1998-02-25 2.0 MYOGLOBIN (HORSE HEART) RECOMBINANT WILD-TYPE COMPLEXED WITH AZIDE
1MNI 1995-04-20 2.07 ALTERATION OF AXIAL COORDINATION BY PROTEIN ENGINEERING IN MYOGLOBIN. BIS-IMIDAZOLE LIGATION IN THE HIS64-->VAL(SLASH)VAL68-->HIS DOUBLE MUTANT
1CRC 1996-03-08 2.08 CYTOCHROME C AT LOW IONIC STRENGTH
3NBT 2010-07-14 2.1 Crystal structure of trimeric cytochrome c from horse heart
1YCB 1994-01-31 2.1 DISTAL POCKET POLARITY IN LIGAND BINDING TO MYOGLOBIN: DEOXY AND CARBONMONOXY FORMS OF A THREONINE68 (E11) MUTANT INVESTIGATED BY X-RAY CRYSTALLOGRAPHY AND INFRARED SPECTROSCOPY
4NFG 2014-09-24 2.11 K13R mutant of horse cytochrome c and yeast cytochrome c peroxidase complex
2IN4 2006-10-31 2.15 Crystal Structure of Myoglobin with Charge Neutralized Heme, ZnDMb-dme
4RSZ 2015-01-14 2.19 The X-ray structure of the Primary Adduct formed in the Reaction between Cisplatin and Cytochrome c
3WI8 2014-03-26 2.2 Crystal structure of horse heart myoglobin reconstituted with manganese porphycene
1MNJ 1995-04-20 2.2 INTERACTIONS AMONG RESIDUES CD3, E7, E10 AND E11 IN MYOGLOBINS: ATTEMPTS TO SIMULATE THE O2 AND CO BINDING PROPERTIES OF APLYSIA MYOGLOBIN
3NBS 2010-07-14 2.2 Crystal structure of dimeric cytochrome c from horse heart
1MNK 1995-04-20 2.2 INTERACTIONS AMONG RESIDUES CD3, E7, E10 AND E11 IN MYOGLOBINS: ATTEMPTS TO SIMULATE THE O2 AND CO BINDING PROPERTIES OF APLYSIA MYOGLOBIN
1MNH 1995-05-08 2.3 INTERACTIONS AMONG RESIDUES CD3, E7, E10 AND E11 IN MYOGLOBINS: ATTEMPTS TO SIMULATE THE O2 AND CO BINDING PROPERTIES OF APLYSIA MYOGLOBIN
5YCG 2018-09-19 2.4 Ancestral myoglobin aMbWp of Pakicetus relative
1KTD 2002-05-01 2.4 CRYSTAL STRUCTURE OF CLASS II MHC MOLECULE IEK BOUND TO PIGEON CYTOCHROME C PEPTIDE
1PMB 1990-01-15 2.5 THE DETERMINATION OF THE CRYSTAL STRUCTURE OF RECOMBINANT PIG MYOGLOBIN BY MOLECULAR REPLACEMENT AND ITS REFINEMENT
1U75 2004-09-28 2.55 Electron Transfer Complex between Horse Heart Cytochrome c and Zinc-Porphyrin Substituted Cytochrome c Peroxidase
2PCB 1993-07-15 2.8 CRYSTAL STRUCTURE OF A COMPLEX BETWEEN ELECTRON TRANSFER PARTNERS, CYTOCHROME C PEROXIDASE AND CYTOCHROME C
1YCA 1994-01-31 2.9 DISTAL POCKET POLARITY IN LIGAND BINDING TO MYOGLOBIN: DEOXY AND CARBONMONOXY FORMS OF A THREONINE68 (E11) MUTANT INVESTIGATED BY X-RAY CRYSTALLOGRAPHY AND INFRARED SPECTROSCOPY
3JBT 2015-11-18 3.8 Atomic structure of the Apaf-1 apoptosome
5JUY 2016-10-19 4.1 Active human apoptosome with procaspase-9
5WVE 2017-02-08 4.4 Apaf-1-Caspase-9 holoenzyme
3J2T 2013-04-10 9.5 An improved model of the human apoptosome

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
91.3 Cytochrome c P00021 CYC_COLLI
92.2 Cytochrome c P00020 CYC_ANAPL
92.3 Cytochrome c P81280 CYC_ALLMI
90.5 Cytochrome c Q52V10 CYC_SAISC
93.3 Cytochrome c P00012 CYC_MIRLE
93.3 Cytochrome c Q52V09 CYC_CEPBA
93.3 Cytochrome c P00013 CYC_MINSC
93.3 Cytochrome c P00014 CYC_MACGI
94.3 Cytochrome c P00011 CYC_CANLF
94.3 Cytochrome c P62898 CYC_RAT
94.3 Cytochrome c P00008 CYC_RABIT
94.3 Cytochrome c P62897 CYC_MOUSE
95.2 Cytochrome c P68098 CYC_LAMGU
95.2 Cytochrome c P68100 CYC_ESCRO
95.2 Cytochrome c P68099 CYC_CAMDR
94.3 Cytochrome c P00007 CYC_HIPAM
97.1 Cytochrome c P62896 CYC_SHEEP
97.1 Cytochrome c P62895 CYC_PIG
97.1 Cytochrome c P62894 CYC_BOVIN
99.0 Cytochrome c P68096 CYC_EQUBU
99.0 Cytochrome c P68097 CYC_EQUAS
100.0 Cytochrome c P00004 CYC_HORSE
90.3 Cytochrome c B4USV4 CYC_OTOGA
90.3 Myoglobin P02163 MYG_ROUAE
90.9 Myoglobin P11343 MYG_LUTLU
90.3 Myoglobin P02165 MYG_TUPGL
90.3 Myoglobin P02167 MYG_NYCCO
90.3 Myoglobin Q0KIY0 MYG_MESST
90.3 Myoglobin P02183 MYG_MESCA
90.8 Myoglobin P02177 MYG_ESCRO
90.8 Myoglobin Q0KIY2 MYG_BALED
90.8 Myoglobin Q0KIY1 MYG_BALBO
90.9 Myoglobin P02189 MYG_PIG
90.9 Myoglobin P02166 MYG_PERPO
92.2 Myoglobin P02169 MYG_LEPMU
91.6 Myoglobin P02181 MYG_INIGE
100.0 Myoglobin P68082 MYG_HORSE
100.0 Myoglobin P68083 MYG_EQUBU
90.1 Myoglobin P02178 MYG_MEGNO