Reversible unfolding of the severe acute respiratory syndrome coronavirus main protease in guanidinium chloride.


Abstract

Chemical denaturant sensitivity of the dimeric main protease from severe acute respiratory syndrome (SARS) coronavirus to guanidinium chloride was examined in terms of fluorescence spectroscopy, circular dichroism, analytical ultracentrifuge, and enzyme activity change. The dimeric enzyme dissociated at guanidinium chloride concentration of <0.4 M, at which the enzymatic activity loss showed close correlation with the subunit dissociation. Further increase in guanidinium chloride induced a reversible biphasic unfolding of the enzyme. The unfolding of the C-terminal domain-truncated enzyme, on the other hand, followed a monophasic unfolding curve. Different mutants of the full-length protease (W31 and W207/W218), with tryptophanyl residue(s) mutated to phenylalanine at the C-terminal or N-terminal domain, respectively, were constructed. Unfolding curves of these mutants were monophasic but corresponded to the first and second phases of the protease, respectively. The unfolding intermediate of the protease thus represented a folded C-terminal domain but an unfolded N-terminal domain, which is enzymatically inactive due to loss of regulatory properties. The various enzyme forms were characterized in terms of hydrophobicity and size-and-shape distributions. We provide direct evidence for the functional role of C-terminal domain in stabilization of the catalytic N-terminal domain of SARS coronavirus main protease. Study holds ProTherm entries: 23604, 23605, 23606, 23607, 23608 Extra Details: Native to intermediate enzymatic activity; unfolding intermediate; hydrophobicity SARS coronavirus

Submission Details

ID: DVi8eLjA4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:54 p.m.

Version: 1

Publication Details
Chang HP;Chou CY;Chang GG,Biophys. J. (2007) Reversible unfolding of the severe acute respiratory syndrome coronavirus main protease in guanidinium chloride. PMID:17142288
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
2HSX 2007-02-06 NMR Structure of the nonstructural protein 1 (nsp1) from the SARS coronavirus
2JZF 2008-02-05 NMR Conformer closest to the mean coordinates of the domain 513-651 of the SARS-CoV nonstructural protein nsp3
2JZD 2008-02-05 NMR structure of the domain 527-651 of the SARS-CoV nonstructural protein nsp3
2RNK 2008-02-05 NMR structure of the domain 513-651 of the SARS-CoV nonstructural protein nsp3
2KQW 2010-02-02 SARS coronavirus-unique domain (SUD): Three-domain molecular architecture in solution and RNA binding. II: Structure of the SUD-C domain of SUD-MC
2KAF 2008-11-25 Solution structure of the SARS-unique domain-C from the nonstructural protein 3 (nsp3) of the severe acute respiratory syndrome coronavirus
2KYS 2010-06-16 NMR Structure of the SARS Coronavirus Nonstructural Protein Nsp7 in Solution at pH 6.5
2GRI 2006-12-19 NMR Structure of the SARS-CoV non-structural protein nsp3a
2GDT 2007-02-06 NMR Structure of the nonstructural protein 1 (nsp1) from the SARS coronavirus
2KQV 2009-12-22 SARS coronavirus-unique domain (SUD): Three-domain molecular architecture in solution and RNA binding. I: Structure of the SUD-M domain of SUD-MC
2LIZ 2012-09-05 NMR solution structure of C-terminal domain of SARS-CoV main protease in 2.5M urea
2IDY 2006-12-05 NMR Structure of the SARS-CoV non-structural protein nsp3a
1YSY 2005-12-06 NMR Structure of the nonstructural Protein 7 (nsP7) from the SARS CoronaVirus
2JZE 2008-02-05 NMR structure of the domain 527-651 of the SARS-CoV nonstructural protein nsp3, single conformer closest to the mean coordinates of an ensemble of twenty energy minimized conformers
2K87 2008-09-16 NMR STRUCTURE OF A PUTATIVE RNA BINDING PROTEIN (SARS1) FROM SARS CORONAVIRUS
2ACF 2006-02-14 1.4 NMR STRUCTURE OF SARS-COV NON-STRUCTURAL PROTEIN NSP3A (SARS1) FROM SARS CORONAVIRUS
4M0W 2014-02-12 1.4 Crystal Structure of SARS-CoV papain-like protease C112S mutant in complex with ubiquitin
5C5O 2016-06-22 1.5 Structure of SARS-3CL protease complex with a phenyl-beta-alanyl (S,R)-N-decalin type inhibitor
3TNT 2012-09-05 1.59 Crystal structure of SARS coronavirus main protease complexed with an alpha, beta-unsaturated ethyl ester inhibitor SG85
4MDS 2013-10-02 1.6 Discovery of N-(benzo[1,2,3]triazol-1-yl)-N-(benzyl)acetamido)phenyl) carboxamides as severe acute respiratory syndrome coronavirus (SARS-CoV) 3CLpro inhibitors: identification of ML300 and non-covalent nanomolar inhibitors with an induced-fit binding
4TWY 2015-02-18 1.6 Structure of SARS-3CL protease complex with a phenylbenzoyl (S,R)-N-decalin type inhibitor
2H2Z 2007-04-03 1.6 Crystal structure of SARS-CoV main protease with authentic N and C-termini
5N19 2018-02-28 1.62 Structure of SARS coronavirus main protease in complex with the alpha-ketoamide (S)-N-benzyl-3-((S)-2-cinnamamido-3-phenylpropanamido)-2-oxo-4-((S)-2-oxopyrrolidin-3-yl)butanamide
5Y3Q 2018-01-10 1.65 Crystal structure of SARS coronavirus papain-like protease conjugated with beta-mercaptoethanol
5Y3E 2018-01-10 1.65 Crystal structure of SARS coronavirus papain-like protease in complex with glycerol
2ZU5 2009-01-13 1.65 complex structure of SARS-CoV 3CL protease with TG-0205486
5C5N 2016-06-22 1.69 Structure of SARS-3CL protease complex with a phenyl-beta-alanyl (R,S)-N-decalin type inhibitor
3SZN 2012-07-25 1.69 Crystal structure of SARS coronavirus main protease complexed with an alpha, beta-unsaturated ethyl ester SG75
2DUC 2007-07-24 1.7 Crystal structure of SARS coronavirus main proteinase(3CLPRO)
2QCY 2008-03-11 1.75 Crystal Structure of a monomeric form of Severe Acute Respiratory Syndrome (SARS) 3C-like protease mutant
2Z94 2007-12-25 1.78 Complex structure of SARS-CoV 3C-like protease with TDT
2Z3C 2007-07-03 1.79 A Mechanistic view of Enzyme Inhibition and Peptide Hydrolysis in the Active Site of the SARS-CoV 3C-Like peptidase
2FYG 2006-08-08 1.8 Crystal structure of NSP10 from Sars coronavirus
2FE8 2006-03-21 1.85 SARS coronavirus papain-like protease: structure of a viral deubiquitinating enzyme
2Z9K 2007-12-25 1.85 Complex structure of SARS-CoV 3C-like protease with JMF1600
2AMD 2005-09-13 1.85 Crystal Structure Of SARS_CoV Mpro in Complex with an Inhibitor N9
1Q2W 2003-07-29 1.86 X-Ray Crystal Structure of the SARS Coronavirus Main Protease
2Z9G 2007-12-25 1.86 Complex structure of SARS-CoV 3C-like protease with PMA
2A5I 2005-10-25 1.88 Crystal structures of SARS coronavirus main peptidase inhibited by an aza-peptide epoxide in the space group C2
3SND 2011-09-07 1.89 Crystal structure of SARS coronavirus main protease complexed with Ac-ESTLQ-H (cocrystallization)
4WY3 2015-02-18 1.89 Structure of SARS-3CL protease complex with a phenylbenzoyl (R,S)-N-decalin type inhibitor
2C3S 2005-10-18 1.9 Structure Of Sars Cov Main Proteinase At 1.9 A (Ph6.5)
2QIQ 2008-02-12 1.9 Structure-based Design and Synthesis and Biological Evaluation of Peptidomimetic SARS-3CLpro Inhibitors
2ALV 2006-08-08 1.9 X-ray structural analysis of SARS coronavirus 3CL proteinase in complex with designed anti-viral inhibitors
1UJ1 2003-11-18 1.9 Crystal structure of SARS Coronavirus Main Proteinase (3CLpro)
2GX4 2007-05-08 1.93 Crystal structure of SARS coronavirus 3CL protease inhibitor complex
2ZU4 2009-01-13 1.93 Complex structure of SARS-CoV 3CL protease with TG-0204998
3VB5 2012-12-12 1.95 Crystal structure of SARS-CoV 3C-like protease with C4Z
2Z9J 2007-12-25 1.95 Complex structure of SARS-CoV 3C-like protease with EPDTC
2HOB 2007-04-03 1.95 Crystal structure of SARS-CoV main protease with authentic N and C-termini in complex with a Michael acceptor N3
3VB7 2012-12-12 1.95 Crystal structure of SARS-CoV 3C-like protease with M4Z
3V3M 2013-01-16 1.96 Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) 3CL Protease in Complex with N-[(1R)-2-(tert-butylamino)-2-oxo-1-(pyridin-3-yl)ethyl]-N-(4-tert-butylphenyl)furan-2-carboxamide inhibitor.
2V6N 2008-07-01 1.98 Crystal structures of the SARS-coronavirus main proteinase inactivated by benzotriazole compounds
3TNS 2012-09-05 1.99 Crystal structure of SARS coronavirus main protease complexed with an alpha, beta-unsaturated ethyl ester inhibitor SG83
3TIT 2012-08-22 1.99 Crystal structure of SARS coronavirus main protease complexed with an alpha, beta-unsaturated ethyl ester SG81
3SN8 2011-09-07 1.99 Crystal structure of SARS coronavirus main protease complexed with Cm-FF-H (soaking)
2BX3 2005-09-26 2.0 Crystal Structure of SARS Coronavirus Main Proteinase (P43212)
3R24 2011-10-26 2.0 Crystal structure of nsp10/nsp16 complex of SARS coronavirus' if possible
3AW1 2011-12-14 2.0 Structure of SARS 3CL protease auto-proteolysis resistant mutant in the absent of inhibitor
5N5O 2018-03-07 2.0 Structure of SARS coronavirus main protease in complex with the alpha-ketoamide (S)-N-benzyl-3-((S)-2-cinnamamido-3-cyclopropylpropanamido)-2-oxo-4-((S)-2-oxopyrrolidin-3-yl)butanamide (Cinnamoyl-cyclopropylalanine-GlnLactam-CO-CO-NH-benzyl)
1WOF 2005-08-30 2.0 Crystal Structure Of SARS-CoV Mpro in Complex with an Inhibitor N1
2XYQ 2011-10-19 2.0 Crystal structure of the nsp16 nsp10 SARS coronavirus complex
2XYV 2011-10-26 2.06 Crystal structure of the nsp16 nsp10 SARS coronavirus complex
3TIU 2012-08-22 2.08 Crystal structure of SARS coronavirus main protease complexed with an alpha,beta-unsaturated ethyl ester inhibitor SG82
2A5A 2005-10-25 2.08 Crystal structure of unbound SARS coronavirus main peptidase in the space group C2
4HI3 2013-05-01 2.09 Crystal structure of dimeric R298A mutant of SARS coronavirus main protease
2Z3D 2007-07-03 2.1 A Mechanistic view of Enzyme Inhibition and Peptide Hydrolysis in the Active Site of the SARS-CoV 3C-Like peptidase
4OW0 2014-04-23 2.1 X-Ray Structural and Biological Evaluation of a Series of Potent and Highly Selective Inhibitors of Human Coronavirus Papain-Like Proteases
2Z9L 2007-12-25 2.1 complex structure of SARS-CoV 3C-like protease with JMF1586
2G9T 2006-08-15 2.1 Crystal structure of the SARS coronavirus nsp10 at 2.1A
5F22 2016-01-27 2.15 C-terminal domain of SARS-CoV nsp8 complex with nsp7
3VB4 2012-12-12 2.2 Crystal structure of SARS-CoV 3C-like protease with B4Z
3VB3 2012-12-12 2.2 Crystal structure of SARS-CoV 3C-like protease in apo form
1UK2 2003-11-18 2.2 Crystal structure of SARS Coronavirus Main Proteinase (3CLpro) At pH8.0
5B6O 2016-06-15 2.2 Crystal structure of MS8104
2W2G 2009-05-26 2.22 Human SARS coronavirus unique domain
3EA8 2009-09-01 2.25 Crystal structure of SARS-CoV main protease triple mutant STI/A in space group C2
3F9F 2009-09-29 2.3 Crystal Structure of the F140A mutant of SARS-Coronovirus 3C-like Protease at pH 6.0
3M3S 2011-03-23 2.3 Crystal structure of SARS-COV main protease Asn214Ala mutant with authorize N-terminus
2AMQ 2005-09-13 2.3 Crystal Structure Of SARS_CoV Mpro in Complex with an Inhibitor N3
3AW0 2011-12-14 2.3 Structure of SARS 3CL protease with peptidic aldehyde inhibitor
2A5K 2005-10-25 2.3 Crystal structures of SARS coronavirus main peptidase inhibited by an aza-peptide epoxide in space group P212121
2Z3E 2007-07-03 2.32 A Mechanistic view of Enzyme Inhibition and Peptide Hydrolysis in the Active Site of the SARS-CoV 3C-Like peptidase
3FZD 2010-05-05 2.35 Mutation of Asn28 disrupts the enzymatic activity and dimerization of SARS 3CLpro
3ATW 2011-12-14 2.36 Structure-Based Design, Synthesis, Evaluation of Peptide-mimetic SARS 3CL Protease Inhibitors
4ZUH 2016-06-15 2.39 Complex structure of PEDV 3CLpro mutant (C144A) with a peptide substrate.
3EA9 2009-09-01 2.4 Crystal structure of SARS-CoV main protease quadruple mutant STIF/A with one molecule in one asymmetric unit
2AHM 2005-11-15 2.4 Crystal structure of SARS-CoV super complex of non-structural proteins: the hexadecamer
1UK3 2003-11-18 2.4 Crystal structure of SARS Coronavirus Main Proteinase (3CLpro) At pH7.6
3SNB 2011-09-07 2.4 Crystal structure of SARS coronavirus main protease complexed with Ac-DSFDQ-H (soaking)
3EBN 2009-05-19 2.4 A Special Dimerization of SARS-CoV Main Protease C-Terminal Domain Due to Domain-swapping
4TWW 2015-02-18 2.42 Structure of SARS-3CL protease complex with a Bromobenzoyl (S,R)-N-decalin type inhibitor
5TL7 2017-05-03 2.44 Crystal structure of SARS-CoV papain-like protease in complex with C-terminal domain mouse ISG15
3AVZ 2011-12-14 2.46 Structure of SARS 3CL protease with peptidic aldehyde inhibitor containing cyclohexyl side chain
3E9S 2008-10-07 2.5 A new class of papain-like protease/deubiquitinase inhibitors blocks SARS virus replication
2XYR 2011-10-26 2.5 Crystal structure of the nsp16 nsp10 SARS coronavirus complex
1UK4 2003-11-18 2.5 Crystal structure of SARS Coronavirus Main Proteinase (3CLpro) Complexed With An Inhibitor
3F9E 2009-09-29 2.5 Crystal Structure of the S139A mutant of SARS-Coronovirus 3C-like Protease
3VB6 2012-12-12 2.5 Crystal structure of SARS-CoV 3C-like protease with C6Z
2PWX 2007-10-30 2.5 Crystal structure of G11A mutant of SARS-CoV 3C-like protease
4OVZ 2014-04-23 2.5 X-Ray Structural and Biological Evaluation of a Series of Potent and Highly Selective Inhibitors of Human Coronavirus Papain-Like Proteases
2Q6G 2008-02-12 2.5 Crystal structure of SARS-CoV main protease H41A mutant in complex with an N-terminal substrate
3E91 2009-08-25 2.55 Crystal structure of SARS-CoV Mpro mutant in P21 at pH6.9
3SNC 2011-09-07 2.58 Crystal structure of SARS coronavirus main protease complexed with Ac-NSTSQ-H (soaking)
3EE7 2009-03-24 2.6 Crystal Structure of SARS-CoV nsp9 G104E
2H85 2006-08-15 2.6 Crystal Structure of Nsp 15 from SARS
3SNE 2011-09-07 2.6 Crystal structure of SARS coronavirus main protease complexed with Ac-ESTLQ-H (Soaking)
3F9G 2009-09-29 2.6 Crystal Structure of the F140A mutant of SARS-Coronovirus 3C-like Protease at pH 6.5
5TL6 2017-05-03 2.62 Crystal structure of SARS-CoV papain-like protease in complex with the C-terminal domain of human ISG15
3MJ5 2010-06-30 2.63 Severe Acute Respiratory Syndrome-Coronavirus Papain-Like Protease Inhibitors: Design, Synthesis, Protein-Ligand X-ray Structure and Biological Evaluation
3EA7 2009-09-01 2.65 Crystal structure of SARS-CoV main protease triple mutant STI/A in space group P21
2QC2 2008-07-01 2.7 Crystal structure of Severe Acute Respiratory Syndrome (SARS) 3C-like protease Asn214Ala mutant
3M3V 2011-03-23 2.7 SARS-CoV main protease triple mutant STI/A with two N-terminal additional residue (Gly-Ser)
1QZ8 2004-02-24 2.7 Crystal structure of SARS coronavirus NSP9
2D2D 2005-09-20 2.7 Crystal Structure Of SARS-CoV Mpro in Complex with an Inhibitor I2
2GA6 2006-08-15 2.7 The crystal structure of SARS nsp10 without zinc ion as additive
3EAJ 2009-09-01 2.7 Crystal structure of SARS-CoV main protease quadruple mutant STIF/A with two molecules in one asymmetric unit
3D62 2008-07-01 2.7 Development of Broad-Spectrum Halomethyl Ketone Inhibitors Against Coronavirus Main Protease 3CLpro
4MM3 2014-07-02 2.75 Crystal structure of SARS-CoV papain-like protease PLpro in complex with ubiquitin aldehyde
2WCT 2009-05-26 2.79 human SARS coronavirus unique domain (triclinic form)
2BX4 2005-09-26 2.79 Crystal Structure of SARS Coronavirus Main Proteinase (P21212)
1UW7 2004-02-20 2.8 Nsp9 protein from SARS-coronavirus.
2RHB 2007-11-27 2.8 Crystal structure of Nsp15-H234A mutant- Hexamer in asymmetric unit
1Z1J 2005-11-22 2.8 Crystal structure of SARS 3CLpro C145A mutant
1Z1I 2005-11-22 2.8 Crystal structure of native SARS CLpro
5E6J 2016-05-18 2.85 Structure of SARS PLpro bound to a Lys48-linked di-ubiquitin activity based probe
2OZK 2007-05-29 2.9 Structure of an N-Terminal Truncated Form of Nendou (NSP15) From SARS-CORONAVIRUS
3M3T 2011-03-23 2.9 SARS-CoV main protease monomeric Arg298Ala mutant with N-terminal additional residues (Gly-Ser)
3F9H 2009-09-29 2.9 Crystal Structure of the F140A mutant of SARS-Coronovirus 3C-like Protease at pH 7.6
3SNA 2011-09-07 3.05 Crystal structure of SARS coronavirus main protease complexed with Ac-NSFSQ-H (soaking)
6NUR 2019-05-29 3.1 SARS-Coronavirus NSP12 bound to NSP7 and NSP8 co-factors
3IWM 2010-07-21 3.2 The octameric SARS-CoV main protease
5C8T 2015-07-15 3.2 Crystal structure of the SARS coronavirus nsp14-nsp10 complex with functional ligand SAM
5C8S 2015-07-15 3.33 Crystal structure of the SARS coronavirus nsp14-nsp10 complex with functional ligands SAH and GpppA
5NFY 2018-01-10 3.38 SARS-CoV nsp10/nsp14 dynamic complex
5C8U 2015-07-15 3.4 Crystal structure of the SARS coronavirus nsp14-nsp10 complex
6NUS 2019-05-29 3.5 SARS-Coronavirus NSP12 bound to NSP8 co-factor

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
99.7 Replicase polyprotein 1ab P0C6F5 R1A_BC279
99.3 Replicase polyprotein 1ab P0C6F8 R1A_BCHK3
99.7 Replicase polyprotein 1ab P0C6T7 R1A_BCRP3
100.0 Replicase polyprotein 1ab P0C6U8 R1A_CVHSA
99.7 Replicase polyprotein 1ab P0C6V9 R1AB_BC279
99.3 Replicase polyprotein 1ab P0C6W2 R1AB_BCHK3
99.7 Replicase polyprotein 1ab P0C6W6 R1AB_BCRP3
100.0 Replicase polyprotein 1ab P0C6X7 R1AB_CVHSA