Effects of salts of alkali earth metals and calcium chloride on the stability of cytochrome c and myoglobin.


Abstract

This study suggests a procedure by which binding of denaturants could be detected without any additional information other than that provided in the denaturation profiles of proteins. Two predominantly alpha-proteins, namely ferricytochrome c and metmyoglobin were denatured by guanidine hydrochloride (GdnHCl) in the presence of low fixed concentrations of salts at 25 degrees C and transition between native and denatured states was followed by absorbance measurements in the visible region (500-350 nm). The raw data were converted into transition curves from which Cm, the midpoint of GdnHCl-induced transition, and deltaGapp, the free energy changes on denaturation, were calculated assuming a two-state mechanism, and values of deltaGapp at zero concentration of the denaturant were estimated. It has been observed (1) that chlorides of Na, K, Cs, and Rb do not affect the native conformation of proteins, (2) that GdnHCl-induced denaturations of proteins in presence and absence of sodium bromide, sodium perchlorate and salts of lithium and calcium are reversible, (3) that optical properties of the GdnHCl-denatured state of proteins remain unchanged in presence of the second denaturant, (4) Cm decreases with an increase in the denaturant concentration, and (5) that except for GdnHCl there exist one or more binding sites on the native proteins for the denaturants. Study holds ProTherm entries: 10028, 10029 Extra Details: binding of denaturants;free changes on denaturation;,two-state mechanism;binding sites

Submission Details

ID: f3iFd2QK3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:39 p.m.

Version: 1

Publication Details
Ahmad Z;Yadav S;Ahmad F;Khan NZ,Biochim. Biophys. Acta (1996) Effects of salts of alkali earth metals and calcium chloride on the stability of cytochrome c and myoglobin. PMID:8639715
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
1OCD 1997-06-16 CYTOCHROME C (OXIDIZED) FROM EQUUS CABALLUS, NMR, MINIMIZED AVERAGE STRUCTURE
1M60 2002-08-07 Solution Structure of Zinc-substituted cytochrome c
1AKK 1997-09-17 SOLUTION STRUCTURE OF OXIDIZED HORSE HEART CYTOCHROME C, NMR, MINIMIZED AVERAGE STRUCTURE
1FI9 2000-08-23 SOLUTION STRUCTURE OF THE IMIDAZOLE COMPLEX OF CYTOCHROME C
2FRC 1997-07-29 CYTOCHROME C (REDUCED) FROM EQUUS CABALLUS, NMR, MINIMIZED AVERAGE STRUCTURE
2GIW 1998-12-09 SOLUTION STRUCTURE OF REDUCED HORSE HEART CYTOCHROME C, NMR, 40 STRUCTURES
1LC1 2003-06-03 Solution Structure Of Reduced Horse Heart Cytochrome c in 30% Acetonitrile Solution, NMR Minimized Average Structure
2N3B 2015-10-28 Structure of oxidized horse heart cytochrome c encapsulated in reverse micelles
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
1GIW 1998-12-09 SOLUTION STRUCTURE OF REDUCED HORSE HEART CYTOCHROME C, NMR, MINIMIZED AVERAGE STRUCTURE
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
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
2V1I 2007-06-12 1.2 Crystal structure of radiation-induced metmyoglobin - aqua ferrous myoglobin at pH 6.8
2FRF 2006-05-30 1.2 Horse Heart Myoglobin, Nitrite Adduct, Crystal Soak
2V1F 2007-06-12 1.2 Crystal structure of radiation-induced myoglobin compound II - intermediate H at pH 8.7
5AZR 2016-01-20 1.2 Crystal structure of aqua-cobalt(III) tetradehydrocorrin in the heme pocket of horse heart myoglobin
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
2V1H 2007-06-12 1.3 Crystal structure of radiation-induced metmyoglobin - aqua ferrous myoglobin at pH 5.2
3WFT 2014-12-03 1.3 Crystal structure of horse heart myoglobin reconstituted with cobalt(II) tetradehydrocorrin
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
2FRK 2006-05-30 1.3 Nitrosyl Horse Heart Myoglobin, Nitric Oxide Gas Method
5DF5 2016-09-14 1.3 The structure of oxidized rat cytochrome c (T28E) at 1.30 angstroms resolution.
2VLX 2009-02-10 1.3 Crystal structure of peroxymyoglobin generated by cryoradiolytic reduction of myoglobin compound III
1GJN 2002-03-01 1.35 Hydrogen Peroxide Derived Myoglobin Compound II at pH 5.2
2V1G 2007-06-12 1.35 Crystal structure of radiation-induced myoglobin compound II - intermediate H 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.
2V1J 2007-06-12 1.4 Crystal structure of radiation-induced metmyoglobin - aqua ferrous myoglobin at pH 8.7
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
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
2VLZ 2008-01-29 1.5 Crystal structure of peroxymyoglobin generated by cryoradiolytic reduction of myoglobin compound III
2B4Z 2005-10-11 1.5 Crystal structure of cytochrome C from bovine heart at 1.5 A resolution.
5YL3 2017-12-27 1.5 Crystal structure of horse heart myoglobin reconstituted with manganese porphycene in resting state at pH 8.5
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.
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
2O5S 2007-10-16 1.6 Cobalt horse heart myoglobin, nitrite modified
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
2O5O 2007-10-16 1.6 Manganese horse heart myoglobin, nitrite modified
5D5R 2015-09-16 1.6 Horse-heart myoglobin - deoxy state
3LR7 2010-07-07 1.6 Ferric horse heart myoglobin, nitrite adduct
2O5T 2007-10-16 1.6 Cobalt horse heart myoglobin, oxidized
1NZ3 2003-04-08 1.6 K45E-K63E Variant of Horse Heart Myoglobin
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
3V2Z 2012-11-21 1.65 Nitrite Bound Chlorin Substituted Myoglobin- Method 2
3V2V 2012-11-21 1.65 Nitrite Bound Chlorin Substituted Myoglobin- Method 1
3VAU 2012-04-11 1.7 Myoglobin nitrite structure: nitriheme modified
1RSE 1996-12-23 1.7 MYOGLOBIN (HORSE HEART) MUTANT WITH SER 92 REPLACED BY ASP (S92D)
1HRM 1995-01-26 1.7 THE PROXIMAL LIGAND VARIANT HIS93TYR OF HORSE HEART MYOGLOBIN
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
1NZ5 2003-04-08 1.7 The Horse heart myoglobin variant K45E/K63E complexed with Manganese
1WLA 1998-01-14 1.7 MYOGLOBIN (HORSE HEART) RECOMBINANT WILD-TYPE
1MWC 1998-08-19 1.7 WILD TYPE MYOGLOBIN WITH CO
1NPG 2003-11-11 1.7 MYOGLOBIN (HORSE HEART) WILD-TYPE COMPLEXED WITH NITROSOETHANE
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
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
5CNE 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 10 ps time delay
5CNC 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.6 ps time delay
5CNB 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.5 ps time delay
5CNG 2015-09-16 1.8 ultrafast dynamics in myoglobin: 150 ps time delay
5CNF 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 50 ps time delay
1MWD 1998-08-19 1.8 WILD TYPE DEOXY MYOGLOBIN
1WEJ 1998-12-09 1.8 IGG1 FAB FRAGMENT (OF E8 ANTIBODY) COMPLEXED WITH HORSE CYTOCHROME C AT 1.8 A RESOLUTION
1BJE 1998-01-28 1.8 H64T VARIANT OF MYOGLOBIN (HORSE HEART) RECOMBINANT WILD-TYPE COMPLEXED WITH AZIDE
3BA2 2008-11-11 1.8 Cyanide bound Chlorin substituted Myoglobin
5CND 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 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
5CMV 2015-09-16 1.8 Ultrafast dynamics in myoglobin: dark-state, CO-ligated structure
5CN8 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.3 ps time delay
3WC8 2013-12-11 1.8 Dimeric horse cytochrome c obtained by refolding with desalting method
5CN5 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0 ps time delay
5CN6 2015-09-16 1.8 Ultrafast dynamics in myoglobin: 0.1 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
5Z7E 2019-01-30 1.8 Horse Heart Myoglobin Mutant - H93M
2O5Q 2007-10-16 1.9 Manganese horse heart myoglobin, nitric oxide modified
1M6C 1998-08-19 1.9 V68N MYOGLOBIN WITH CO
3O20 2012-01-25 1.9 Electron transfer complexes:experimental mapping of the Redox-dependent Cytochrome C electrostatic surface
1HRC 1994-11-01 1.9 HIGH-RESOLUTION THREE-DIMENSIONAL STRUCTURE OF HORSE HEART CYTOCHROME C
5Z7F 2019-01-30 1.9 Horse Heart Myoglobin Mutant -V68A/I107Y-Sulfide Derivative
2NSR 2007-11-06 1.9 Nitromethane Modified Horse Heart Myoglobin
1NZ2 2003-04-08 1.9 K45E Variant of Horse Heart Myoglobin
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)
1HSY 1995-02-27 1.9 ORIGIN OF THE PH-DEPENDENT SPECTROSCOPIC PROPERTIES OF PENTACOORDINATE METMYOGLOBIN VARIANTS
3HEN 2009-12-29 1.9 Ferric Horse Heart Myoglobin; H64V/V67R Mutant
1YMB 1994-01-31 1.9 HIGH RESOLUTION STUDY OF THE THREE-DIMENSIONAL STRUCTURE OF HORSE HEART METMYOGLOBIN
1NPF 2003-11-11 1.9 MYOGLOBIN (HORSE HEART) WILD-TYPE COMPLEXED WITH NITRIC OXIDE
3RJN 2012-05-09 1.9 Horse heart myoglobin: D44K/D60K mutant with zinc (II) -deuteroporphyrin dimethyl ester
1MYJ 1994-01-31 1.9 DISTAL POLARITY IN LIGAND BINDING TO MYOGLOBIN: STRUCTURAL AND FUNCTIONAL CHARACTERIZATION OF A THREONINE68(E11) MUTANT
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)
3HC9 2009-12-29 2.0 Ferric Horse Heart Myoglobin; H64V mutant
3WYO 2014-11-19 2.0 Heterodimeric myoglobin formed by domain swapping
1YMA 1994-01-31 2.0 STRUCTURAL CHARACTERIZATION OF HEME LIGATION IN THE HIS64-->TYR VARIANT OF MYOGLOBIN
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)
2O5B 2007-10-16 2.0 Manganese horse heart myoglobin, reduced
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
3HEO 2009-12-29 2.0 Ferric Horse Heart Myoglobin; H64V/V67R mutant, Nitrite Modified
1AZI 1998-02-25 2.0 MYOGLOBIN (HORSE HEART) RECOMBINANT WILD-TYPE COMPLEXED WITH AZIDE
1YMC 1994-01-31 2.0 THREE-DIMENSIONAL STRUCTURE OF CYANOMET-SULFMYOGLOBIN C
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
3NBS 2010-07-14 2.2 Crystal structure of dimeric cytochrome c from horse heart
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
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