Contribution of conformational stability and reversibility of unfolding to the increased thermostability of human and bovine superoxide dismutase mutated at free cysteines.


Abstract

The conformational stability and reversibility of unfolding of the human dimeric enzyme Cu Zn superoxide dismutase (HSOD) and the three mutant enzymes constructed by replacement of Cys6 by Ala and Cys111 by Ser, singly and in combination, were determined by differential scanning calorimetry. The differential scanning calorimetry profile of wild-type HSOD consists of two components, which probably represent the unfolding of the oxidized and reduced forms of the enzyme, with denaturation temperatures (Tm) of 74.9 and 83.6 degrees C, approximately 7 degrees lower than those for bovine superoxide dismutase (BSOD). The conformational stabilities of the two components of the mutant HSOD's differ only slightly from those of the wild type (delta delta Gs of -0.2 to +0.8 kcal/mol of dimer), while replacement of the BSOD Cys6 by Ala is somewhat destabilizing (delta delta G of -0.7 to -1.3 kcal/mol of dimer). These small alterations in conformational stability do not correlate with the large increases in resistance to thermal inactivation following substitution of free Cys in both HSOD and BSOD (McRee, D.E., Redford, S.M., Getzoff, E.D., Lepock, J.R., Hallewell, R.A., and Tainer, J.A. (1990) J. Biol. Chem. 265, 14234-14241 and Hallewell, R.A., Imlay, K.C., Laria, I., Gallegos, C., Fong, N., Irvine, B., Getzoff, E.D., Tainer, J.A., Cubelli, D.E., Bielski, B.H.J., Olson, P., Mallenbach, G.T., and Cousens, L.S. (1991) Proteins Struct. Funct. Genet., submitted for publication). The reversibility of unfolding was determined by scanning part way through the profile, cooling, rescanning, and calculating the amount of protein irreversibly unfolded by the first scan. The order of reversibility at a constant level of unfolding is the same as the order of resistance to inactivation for both the HSOD and BSOD wild-type and mutant enzymes. Thus, the greater resistance to thermal inactivation of the superoxide dismutase enzymes with free Cys replaced by Ala or Ser is dominated by a greater resistance to irreversible unfolding and relatively unaffected by changes in conformational stability. Study holds ProTherm entries: 5179, 5180, 5181, 5182, 5183 Extra Details: free cysteines; conformational stability; reversibility;,thermal inactivation

Submission Details

ID: yq6BcGwJ3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:29 p.m.

Version: 1

Publication Details
Lepock JR;Frey HE;Hallewell RA,J. Biol. Chem. (1990) Contribution of conformational stability and reversibility of unfolding to the increased thermostability of human and bovine superoxide dismutase mutated at free cysteines. PMID:2254318
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
1KMG 2002-10-02 The Solution Structure Of Monomeric Copper-free Superoxide Dismutase
2MP3 2015-05-20 Truncated L126Z-sod1 in DPC micelle
1RK7 2003-12-02 Solution structure of apo Cu,Zn Superoxide Dismutase: role of metal ions in protein folding
1DSW 2000-03-22 THE SOLUTION STRUCTURE OF A MONOMERIC, REDUCED FORM OF HUMAN COPPER, ZINC SUPEROXIDE DISMUTASE BEARING THE SAME CHARGE AS THE NATIVE PROTEIN
2NAM 2016-12-14 Full-length WT SOD1 in DPC MICELLE
1BA9 1998-09-16 THE SOLUTION STRUCTURE OF REDUCED MONOMERIC SUPEROXIDE DISMUTASE, NMR, 36 STRUCTURES
2LU5 2012-06-27 Structure and chemical shifts of Cu(I),Zn(II) superoxide dismutase by solid-state NMR
2AF2 2005-11-15 Solution structure of disulfide reduced and copper depleted Human Superoxide Dismutase
1L3N 2002-05-08 The Solution Structure of Reduced Dimeric Copper Zinc SOD: the Structural Effects of Dimerization
4A7U 2012-11-28 0.98 Structure of human I113T SOD1 complexed with adrenaline in the p21 space group.
2WYT 2010-10-27 1.0 1.0 A resolution structure of L38V SOD1 mutant
4A7V 2012-11-28 1.0 Structure of human I113T SOD1 mutant complexed with dopamine in the p21 space group
1MFM 1999-04-21 1.02 MONOMERIC HUMAN SOD MUTANT F50E/G51E/E133Q AT ATOMIC RESOLUTION
4A7S 2012-12-05 1.06 Structure of human I113T SOD1 mutant complexed with 5-Fluorouridine in the p21 space group
2C9V 2005-12-20 1.07 Atomic resolution structure of Cu-Zn Human Superoxide dismutase
2V0A 2007-06-19 1.15 Atomic resolution crystal structure of Human Superoxide Dismutase
1Q0E 2003-09-23 1.15 Atomic resolution (1.15 ) crystal structure of bovine copper, zinc superoxide dismutase
4A7Q 2012-10-24 1.22 Structure of human I113T SOD1 mutant complexed with 4-(4-methyl-1,4- diazepan-1-yl)quinazoline in the p21 space group.
4A7G 2012-10-24 1.24 Structure of human I113T SOD1 mutant complexed with 4-methylpiperazin- 1-yl)quinazoline in the p21 space group.
2C9U 2005-12-16 1.24 1.24 Angstroms resolution structure of as-isolated Cu-Zn Human Superoxide dismutase
2C9S 2005-12-15 1.24 1.24 Angstroms resolution structure of Zn-Zn Human Superoxide dismutase
6Z3V 2020-09-16 1.25 A4V mutant of human SOD1 bound with N-aryl benzoisoselenazolone derivative 13 in P21 space group
5J0F 2017-02-01 1.25 Monomeric Human Cu,Zn Superoxide dismutase, loops IV and VII deleted, apo form, circular permutant P4/5
4BCY 2013-02-27 1.27 Monomeric Human Cu,Zn Superoxide dismutase, mutation H43F
5O3Y 2018-06-13 1.3 SOD1 bound to Ebsulfur
2VR6 2008-04-15 1.3 Crystal Structure of G85R ALS mutant of Human Cu,Zn Superoxide Dismutase (CuZnSOD) at 1.3 A resolution
1OZU 2003-05-27 1.3 Crystal Structure of Familial ALS Mutant S134N of human Cu,Zn Superoxide Dismutase (CuZnSOD) to 1.3A resolution
4NIO 2013-12-04 1.3 GVTGIAQ segment 147-153 from Human Superoxide Dismutase with I149T mutation associated with a familial form of amyotrophic lateral sclerosis
2VR8 2008-04-08 1.36 Crystal Structure of G85R ALS mutant of Human Cu,Zn Superoxide Dismutase (CuZnSOD) at 1.36 A resolution
6Z4O 2020-09-16 1.4 A4V mutant of human SOD1 bound with benzyl benzoisoselenazolone derivative 1 in P21 space group
5WMJ 2018-03-28 1.4 KVWGSI segment from Superoxide Dismutase 1,residues 30-35
4NIN 2013-12-04 1.4 DSVISLS segment 101-107 from Human Superoxide Dismutase
1P1V 2003-08-26 1.4 Crystal Structure of FALS-associated human Copper-Zinc Superoxide Dismutase (CuZnSOD) Mutant D125H to 1.4A
3GZQ 2009-10-13 1.4 HUMAN SOD1 A4V Metal-free Variant
4A7T 2012-11-28 1.45 Structure of human I113T SOD1 mutant complexed with isoproteranol in the p21 space group
6Z4G 2020-09-16 1.45 A4V mutant of human SOD1 bound with ebselen in P21 space group
2ZOW 2009-06-30 1.45 Crystal Structure of H2O2 treated Cu,Zn-SOD
6Z4K 2020-09-16 1.45 A4V mutant of human SOD1 bound with benzyl benzoisoselenazolone derivative 6 in P21 space group
2XJK 2010-09-01 1.45 Monomeric Human Cu,Zn Superoxide dismutase
5O40 2018-06-13 1.5 SOD1 bound to Ebselen
2WZ5 2010-12-08 1.5 L38V SOD1 mutant complexed with L-methionine.
6Z4M 2020-09-16 1.55 A4V mutant of human SOD1 bound with 2-(pyridin-3-ylmethyl)benzoisoselenazolone derivative 10 in P21 space group
6Z4J 2020-09-16 1.55 A4V mutant of human SOD1 bound with benzyl benzoisoselenazolone derivative 5 in P21 space group
3H2P 2010-05-05 1.55 Human SOD1 D124V Variant
2XJL 2010-09-01 1.55 Monomeric Human Cu,Zn Superoxide dismutase without Cu ligands
2Z7Y 2008-09-02 1.55 Crystal Structure of H2O2 treated Cu,Zn-SOD
2WZ6 2010-12-08 1.55 G93A SOD1 mutant complexed with Quinazoline.
2VR7 2008-04-15 1.58 Crystal Structure of G85R ALS mutant of Human Cu,Zn Superoxide Dismutase (CuZnSOD) at 1.58 A resolution
1UXL 2004-03-19 1.6 I113T mutant of human SOD1
6Z4L 2020-09-16 1.6 A4V mutant of human SOD1 bound with 2-(pyridin-3-ylmethyl)benzoisoselenazolone derivative 9 in P21 space group
6Z4I 2020-09-16 1.6 A4V mutant of human SOD1 bound with benzyl benzoisoselenazolone derivative 3 in P21 space group
5J0C 2017-02-01 1.6 Monomeric Human Cu,Zn Superoxide dismutase, loops IV and VII deleted, apo form, circular permutant P2/3
6SPA 2020-03-18 1.65 A4V MUTANT OF HUMAN SUPEROXIDE DISMUTASE 1 IN C2 SPACE GROUP
1CBJ 1999-03-03 1.65 CRYSTAL STRUCTURE OF BOVINE SUPEROXIDE DISMUTASE CRYSTAL.
3QQD 2011-03-09 1.65 Human SOD1 H80R variant, P212121 crystal form
2GBT 2007-01-02 1.7 C6A/C111A CuZn Superoxide dismutase
2WYZ 2010-10-27 1.7 L38V SOD1 mutant complexed with UMP
1E9P 2000-12-03 1.7 Crystal structure of bovine Cu, Zn SOD to 1.7 Angstrom (3 of 3)
1PU0 2003-09-09 1.7 Structure of Human Cu,Zn Superoxide Dismutase
2WZ0 2010-12-08 1.72 L38V SOD1 mutant complexed with aniline.
3K91 2010-01-19 1.75 Polysulfane Bridge in Cu-Zn Superoxide Dismutase
1E9Q 2000-12-03 1.75 Crystal structure of bovine Cu Zn SOD - (1 of 3)
6FLH 2018-11-07 1.79 Monomeric Human Cu,Zn Superoxide dismutase, SOD1 7+7, apo form
3T5W 2012-08-01 1.8 2ME modified human SOD1
1HL5 2003-05-08 1.8 The Structure of Holo Type Human Cu, Zn Superoxide Dismutase
2Z7W 2008-09-02 1.8 Crystal Structure of H2O2 treated Cu,Zn-SOD
2AEO 2006-05-02 1.8 Crystal structure of cisplatinated bovine Cu,Zn superoxide dismutase
1PTZ 2003-09-09 1.8 Crystal structure of the human CU, Zn Superoxide Dismutase, Familial Amyotrophic Lateral Sclerosis (FALS) Mutant H43R
6B79 2018-05-30 1.8 Curved pair of sheets formed from SOD1 residues 28-38 with familial mutation G37R.
1HL4 2003-05-08 1.82 The Structure of Apo Type Human Cu, Zn Superoxide Dismutase
1E9O 2000-12-03 1.85 Crystal structure of bovine SOD - 1 of 3
2Z7Z 2008-09-02 1.85 Crystal Structure of H2O2 treated Cu,Zn-SOD
3H2Q 2010-05-05 1.85 Human SOD1 H80R variant, P21 crystal form
1N19 2002-11-27 1.86 Structure of the HSOD A4V mutant
5YUL 2018-11-21 1.9 Native Structure of hSOD1 in P6322 space group
5YTO 2018-11-21 1.9 Crystal Structure of human Superoxide Dismutase I (hSOD1) in complex with a napthalene-catechol linked compound.
1SXC 1995-06-03 1.9 CRYSTAL STRUCTURE OF REDUCED BOVINE ERYTHROCYTE SUPEROXIDE DISMUTASE AT 1.9 ANGSTROMS RESOLUTION
3ECV 2009-05-19 1.9 Crystal structure of the ALS-related pathological mutant I113T of human apo Cu,Zn Superoxide Dismutase (SOD1)
1SXN 1998-03-18 1.9 REDUCED BOVINE SUPEROXIDE DISMUTASE AT PH 5.0
1AZV 1998-02-25 1.9 FAMILIAL ALS MUTANT G37R CUZNSOD (HUMAN)
1SXA 1995-06-03 1.9 CRYSTAL STRUCTURE OF REDUCED BOVINE ERYTHROCYTE SUPEROXIDE DISMUTASE AT 1.9 ANGSTROMS RESOLUTION
1UXM 2004-03-19 1.9 A4V mutant of human SOD1
3CQQ 2008-04-29 1.9 Human SOD1 G85R Variant, Structure II
2ZKW 2009-03-24 1.9 Crystal structure of human Cu-Zn superoxide dismutase mutant G85R in space group P21
4NIP 2013-12-04 1.9 GVIGIAQ segment 147-153 from Human Superoxide Dismutase
3ECU 2009-05-19 1.9 Crystal structure of human apo Cu,Zn Superoxide Dismutase (SOD1)
5YTU 2018-11-21 1.9 Structure of human SOD1 complexed with isoproteranol in C2221 space group
4BCZ 2013-02-27 1.93 Monomeric Human Cu,Zn Superoxide dismutase, loops IV and VII deleted, apo form.
6FFK 2018-11-28 1.94 Human apo-SOD1 bound to PtCl2(1R,2R-1,4-DACH
6Z4H 2020-09-16 1.95 A4V mutant of human SOD1 bound with benzyl benzoisoselenazolone derivative 2 in P21 space group
3CQP 2008-04-29 1.95 Human SOD1 G85R Variant, Structure I
6SPJ 2020-03-18 1.97 A4V MUTANT OF HUMAN SOD1 WITH EBSELEN DERIVATIVE 1
2WKO 2009-11-24 1.97 Structure of metal loaded Pathogenic SOD1 Mutant G93A.
5K02 2016-11-23 1.99 Structure of human SOD1 with T2D mutation
2GBV 2007-01-02 2.0 C6A/C111A/C57A/C146A holo CuZn Superoxide dismutase
2R27 2007-12-11 2.0 Constitutively zinc-deficient mutant of human superoxide dismutase (SOD), C6A, H80S, H83S, C111S
5IIW 2017-06-28 2.0 Corkscrew assembly of SOD1 residues 28-38 without potassium iodide
1COB 1993-10-31 2.0 CRYSTAL STRUCTURE SOLUTION AND REFINEMENT OF THE SEMISYNTHETIC COBALT SUBSTITUTED BOVINE ERYTHROCYTE ENZYME SUPEROXIDE DISMUTASE AT 2.0 ANGSTROMS RESOLUTION
2SOD 1980-05-07 2.0 DETERMINATION AND ANALYSIS OF THE 2 ANGSTROM STRUCTURE OF COPPER, ZINC SUPEROXIDE DISMUTASE
1SXS 1998-09-30 2.0 Reduced bovine superoxide dismutase at pH 5.0 complexed with thiocyanate
3HW7 2010-06-23 2.0 High pressure (0.57 GPa) crystal structure of bovine copper, zinc superoxide dismutase at 2.0 angstroms
1N18 2002-11-27 2.0 Thermostable mutant of Human Superoxide Dismutase, C6A, C111S
1SXB 1995-06-03 2.0 CRYSTAL STRUCTURE OF REDUCED BOVINE ERYTHROCYTE SUPEROXIDE DISMUTASE AT 1.9 ANGSTROMS RESOLUTION
6FOI 2019-01-30 2.0 Human Cys57/156Ala superoxide dismutase-1 (SOD1), as isolated.
2GBU 2007-01-02 2.0 C6A/C111A/C57A/C146A apo CuZn Superoxide dismutase
6DTK 2019-06-19 2.0 Heterodimers of FALS mutant SOD enzyme
5J07 2017-02-01 2.0 Monomeric Human Cu,Zn Superoxide dismutase, loops IV and VII deleted, apo form, circular permutant P1/2
1SXZ 1998-09-30 2.05 Reduced bovine superoxide dismutase at pH 5.0 complexed with azide
3GZO 2009-10-13 2.1 HUMAN SOD1 G93A Variant
2Z7U 2008-09-02 2.1 Crystal Structure of H2O2 treated Cu,Zn-SOD
3SOD 1993-04-15 2.1 CHANGES IN CRYSTALLOGRAPHIC STRUCTURE AND THERMOSTABILITY OF A CU,ZN SUPEROXIDE DISMUTASE MUTANT RESULTING FROM THE REMOVAL OF BURIED CYSTEINE
5DLI 2016-09-14 2.1 Corkscrew assembly of SOD1 residues 28-38
3ECW 2009-05-19 2.15 Crystal structure of the ALS-related pathological mutant T54R of human apo Cu,Zn Superoxide Dismutase (SOD1)
1OEZ 2003-05-29 2.15 Zn His46Arg mutant of Human Cu, Zn Superoxide Dismutase
4B3E 2012-09-26 2.15 Structure of copper-zinc superoxide dismutase complexed with bicarbonate.
3GTV 2010-09-08 2.2 Human-mouse SOD1 chimera
3HFF 2009-06-16 2.2 Monomeric human Cu,Zn Superoxide dismutase without Zn ligands
4MCM 2014-08-27 2.2 Human SOD1 C57S Mutant, As-isolated
3GQF 2009-04-07 2.2 Structural and Biophysical Properties of the Pathogenic SOD1 Variant H46R/H48Q
6SPH 2020-03-18 2.25 A4V MUTANT OF HUMAN SUPEROXIDE DISMUTASE 1 WITH EBSELEN BOND IN C2 SPACE GROUP
3RE0 2012-04-25 2.28 Crystal structure of human apo Cu,Zn superoxide dismutase (SOD1) complexed with cisplatin
1CB4 1999-03-03 2.3 CRYSTAL STRUCTURE OF COPPER, ZINC SUPEROXIDE DISMUTASE
2NNX 2006-11-07 2.3 Crystal Structure of the H46R, H48Q double mutant of human [Cu-Zn] Superoxide Dismutase
5U9M 2017-05-31 2.35 Copper-Zinc Superoxide Dismutase is Activated through a Sulfenic Acid Intermediate at a Copper-ion Entry Site
4OH2 2014-10-15 2.38 Crystal Structure of Cu/Zn Superoxide Dismutase I149T
1SPD 1994-04-30 2.4 AMYOTROPHIC LATERAL SCLEROSIS AND STRUCTURAL DEFECTS IN CU,ZN SUPEROXIDE DISMUTASE
2ZKY 2009-03-24 2.4 Crystal structure of human Cu-Zn superoxide dismutase mutant G93A
3LTV 2010-09-08 2.45 Mouse-human sod1 chimera
4FF9 2013-09-04 2.5 Crystal Structure of cysteinylated WT SOD1.
5J0G 2017-02-01 2.5 Monomeric Human Cu,Zn Superoxide dismutase, loops IV and VII deleted, apo form, circular permutant P7/8
6A9O 2019-07-17 2.5 Rational discovery of a SOD1 tryptophan oxidation inhibitor with therapeutic potential for amyotrophic lateral sclerosis
1SOS 1993-04-15 2.5 ATOMIC STRUCTURES OF WILD-TYPE AND THERMOSTABLE MUTANT RECOMBINANT HUMAN CU, ZN SUPEROXIDE DISMUTASE
1SDA 1993-10-31 2.5 CRYSTAL STRUCTURE OF PEROXYNITRITE-MODIFIED BOVINE CU,ZN SUPEROXIDE DISMUTASE
1OZT 2003-05-27 2.5 Crystal Structure of apo-H46R Familial ALS Mutant human Cu,Zn Superoxide Dismutase (CuZnSOD) to 2.5A resolution
6FOL 2019-01-30 2.55 Domain II of the human copper chaperone in complex with human Cu,Zn superoxide dismutase
4MCN 2014-08-27 2.6 Human SOD1 C57S Mutant, Metal-free
2ZKX 2009-03-24 2.72 Crystal structure of human Cu-Zn superoxide dismutase mutant G85R in space group I212121
6SPK 2020-03-18 2.77 A4V MUTANT OF HUMAN SOD1 WITH EBSELEN DERIVATIVE 6
5WOR 2018-05-30 2.77 Corkscrew assembly of SOD1 residues 28-38 with familial mutation G37R
4BD4 2013-02-27 2.78 Monomeric Human Cu,Zn Superoxide dismutase, loops IV and VII deleted, apo form, mutant H43F
6SPI 2020-03-18 2.8 A4V MUTANT OF HUMAN SOD1 WITH EBSELEN DERIVATIVE 4
1FUN 1999-07-23 2.85 SUPEROXIDE DISMUTASE MUTANT WITH LYS 136 REPLACED BY GLU, CYS 6 REPLACED BY ALA AND CYS 111 REPLACED BY SER (K136E, C6A, C111S)
6FP6 2019-01-30 3.0 Complex of human Cu,Zn SOD1 with the human copper chaperone for SOD1 in a compact conformation
6FON 2019-01-30 3.05 Elongated conformer of the human copper chaperone for SOD1 complexed with human SOD1
3GZP 2009-10-13 3.1 HUMAN SOD1 G93A Metal-free Variant
3KH4 2010-08-11 3.5 Crystal structure of human Cu/Zn superoxide dismutase recombinantly produced in Leishmania tarantolae; P6522 crystal form containing 6 chains in the asymmetric unit
3KH3 2010-08-11 3.5 Crystal structure of human Cu/Zn superoxide dismutase recombinantly produced in Leishmania tarantolae; P212121 crystal form containing 12 chains in the asymmetric unit
4XCR 2016-01-20 3.6 Monomeric Human Cu,Zn Superoxide dismutase, loops IV and VII deleted, apo form, mutant I35A

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
94.7 Superoxide dismutase [Cu-Zn] O46412 SODC_CEREL
97.4 Superoxide dismutase [Cu-Zn] P09670 SODC_SHEEP
98.0 Superoxide dismutase [Cu-Zn] Q5FB29 SODC_CAPHI
98.7 Superoxide dismutase [Cu-Zn] Q52RN5 SODC_BOSMU
100.0 Superoxide dismutase [Cu-Zn] P00442 SODC_BOVIN
91.6 Superoxide dismutase [Cu-Zn] Q8HXQ0 SODC_MACMU
91.6 Superoxide dismutase [Cu-Zn] Q8HXQ2 SODC_MACFU
91.6 Superoxide dismutase [Cu-Zn] Q8HXQ1 SODC_MACFA
92.2 Superoxide dismutase [Cu-Zn] Q8HXP8 SODC_CALJA
93.5 Superoxide dismutase [Cu-Zn] Q8HXP9 SODC_SAPAP
94.2 Superoxide dismutase [Cu-Zn] Q8HXQ3 SODC_HYLLA
95.5 Superoxide dismutase [Cu-Zn] Q8HXQ4 SODC_PONPY
100.0 Superoxide dismutase [Cu-Zn] P60052 SODC_PANTR
100.0 Superoxide dismutase [Cu-Zn] P00441 SODC_HUMAN