Reversible dissociation and unfolding of the dimeric protein thymidylate synthase.


Abstract

Conditions for in vitro unfolding and refolding of dimeric thymidylate synthase from Lactobacillus casei were found. Ultraviolet difference and circular dichroism spectra showed that the enzyme was completely unfolded at concentrations of urea over 5.5 M. As measured by restoration of enzyme activity, refolding was accomplished when 0.5 M potassium chloride was included in the refolding mixture. Recombination of subunits from catalytically inactive mutant homodimers to form an active hybrid dimer was achieved under these unfolding-refolding conditions, demonstrating a monomer to dimer association step. Study holds ProTherm entries: 10382 Extra Details: additive : EDTA(0.1 mM), dimerization; folding; oligomerization; thymidylate synthase

Submission Details

ID: fw7pNpXU3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:40 p.m.

Version: 1

Publication Details
Perry KM;Pookanjanatavip M;Zhao J;Santi DV;Stroud RM,Protein Sci. (1992) Reversible dissociation and unfolding of the dimeric protein thymidylate synthase. PMID:1304920
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
3IK0 2010-08-11 2.1 Lactobacillus casei Thymidylate Synthase in Ternary Complex with dUMP and the Phtalimidic Derivative 7C1
1NJD 1996-07-11 2.2 THYMIDYLATE SYNTHASE, MUTATION, N229D WITH 2'-DEOXYURIDINE 5'-MONOPHOSPHATE (DUMP)
1TSY 1996-03-08 2.2 THYMIDYLATE SYNTHASE R179K MUTANT
1JMG 1998-01-28 2.2 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
3IJZ 2010-08-11 2.21 Lactobacillus casei Thymidylate Synthase ternary complex with dUMP and Pthalimidic derivative 15C
3IK1 2010-08-11 2.25 Lactobacillus casei Thymidylate Synthase in Ternary Complex with dUMP and the Phtalimidic Derivative 20C
1VZE 1997-03-12 2.3 L. CASEI THYMIDYLATE SYNTHASE MUTANT E60Q TERNARY COMPLEX WITH DUMP AND CB3717
1NJE 1996-07-11 2.3 THYMIDYLATE SYNTHASE WITH 2'-DEOXYCYTIDINE 5'-MONOPHOSPHATE (DCMP)
1TVV 1998-01-28 2.3 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
4TMS 1992-01-15 2.35 PLASTIC ADAPTATION TOWARD MUTATIONS IN PROTEINS: STRUCTURAL COMPARISON OF THYMIDYLATE SYNTHASES
1BPJ 1998-08-19 2.4 THYMIDYLATE SYNTHASE R178T, R179T DOUBLE MUTANT
2G8A 2006-03-14 2.4 Lactobacillus casei Y261M in complex with substrate, dUMP
1BP0 2000-01-12 2.4 THYMIDYLATE SYNTHASE R23I MUTANT
3BYX 2009-01-20 2.4 Lactobacillus CASEI Thymidylate Synthase Ternary Complex with DUMP and the Phtalimidic Derivative C00 in Multiple Binding Modes
1BP6 1998-08-19 2.4 THYMIDYLATE SYNTHASE R23I, R179T DOUBLE MUTANT
1BO8 1998-08-19 2.4 THYMIDYLATE SYNTHASE R178T MUTANT
2G86 2006-03-14 2.4 L. casei thymidylate synthase Y261F in complex with substrate, dUMP
2G8D 2006-03-14 2.4 Lactobacillus casei thymidylate synthase Y261W-dUMP complex
3C0A 2009-01-20 2.4 Lactobacillus CASEI Thymidylate Synthase Ternary Complex with DUMP and the Phtalimidic Derivative 14C in Multiple Binding Modes-Mode 2
1BO7 1998-08-19 2.4 THYMIDYLATE SYNTHASE R179T MUTANT
1TVW 1998-01-28 2.5 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1JMH 1998-01-28 2.5 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1JMI 1998-01-28 2.5 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1TSX 1996-03-08 2.5 THYMIDYLATE SYNTHASE R179E MUTANT
1NJC 1996-07-11 2.5 THYMIDYLATE SYNTHASE, MUTATION, N229D WITH 2'-DEOXYCYTIDINE 5'-MONOPHOSPHATE (DCMP)
1TVU 1998-01-28 2.5 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1VZC 1997-03-12 2.5 L. CASEI THYMIDYLATE SYNTHASE MUTANT E60Q BINARY COMPLEX WITH FDUMP
1LCA 1995-10-15 2.5 LACTOBACILLUS CASEI THYMIDYLATE SYNTHASE TERNARY COMPLEX WITH DUMP AND CB3717
1NJA 1996-07-11 2.5 THYMIDYLATE SYNTHASE, MUTATION, N229C WITH 2'-DEOXYCYTIDINE 5'-MONOPHOSPHATE (DCMP)
1VZB 1997-03-12 2.5 L. CASEI THYMIDYLATE SYNTHASE MUTANT E60Q BINARY COMPLEX WITH DUMP
1JMF 1998-01-28 2.5 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1TSW 1996-03-08 2.5 THYMIDYLATE SYNTHASE R179A MUTANT
1VZA 1997-03-12 2.5 THYMIDYLATE SYNTHASE E60D MUTANT BINARY COMPLEX WITH 2'-DEOXYURIDINE 5'-MONOPHOSPHATE (DUMP)
1LCE 1995-10-15 2.5 LACTOBACILLUS CASEI THYMIDYLATE SYNTHASE TERNARY COMPLEX WITH DUMP AND CH2THF
1LCB 1995-10-15 2.5 LACTOBACILLUS CASEI THYMIDYLATE SYNTHASE TERNARY COMPLEX WITH DTMP AND H2FOLATE
1TSL 1998-06-17 2.5 L. CASEI THYMIDYLATE SYNTHASE WITH SPECIES SPECIFIC INHIBITOR
2G89 2006-03-14 2.5 L. casei thymidylate synthase Y261A in complex with substrate, dUMP
1VZD 1997-03-12 2.5 L. CASEI THYMIDYLATE SYNTHASE MUTANT E60Q TERNARY COMPLEX WITH FDUMP AND CB3717
2TDM 1997-08-20 2.55 STRUCTURE OF THYMIDYLATE SYNTHASE
3BNZ 2008-12-16 2.6 Crystal structure of Thymidylate Synthase ternary complex with dUMP and 8A inhibitor
3C06 2009-01-20 2.6 Lactobacillus CASEI Thymidylate Synthase Ternary Complex With DUMP and the Phtalimidic Derivative 14C in Multiple Binding Modes-Mode 1
1TDB 1993-07-15 2.65 STRUCTURES OF THYMIDYLATE SYNTHASE WITH A C-TERMINAL DELETION: ROLE OF THE C-TERMINUS IN ALIGNMENT OF D/UMP AND CH2H4FOLATE
1TDC 1993-07-15 2.65 STRUCTURES OF THYMIDYLATE SYNTHASE WITH A C-TERMINAL DELETION: ROLE OF THE C-TERMINUS IN ALIGNMENT OF D/UMP AND CH2H4FOLATE
3BZ0 2009-01-20 2.7 Lactobacillus Casei Thymidylate Synthase Ternary Complex with DUMP and the Phtalimidic Derivative C00
2TDD 1993-07-15 2.7 STRUCTURES OF THYMIDYLATE SYNTHASE WITH A C-TERMINAL DELETION: ROLE OF THE C-TERMINUS IN ALIGNMENT OF D/UMP AND CH2H4FOLATE
1NJB 1996-10-14 2.74 THYMIDYLATE SYNTHASE
1TSZ 1996-03-08 2.75 THYMIDYLATE SYNTHASE R179K MUTANT
1TSV 1996-03-08 2.9 THYMIDYLATE SYNTHASE R179A MUTANT
1THY 1993-10-31 2.9 REFINED STRUCTURES OF SUBSTRATE-BOUND AND PHOSPHATE-BOUND THYMIDYLATE SYNTHASE FROM LACTOBACILLUS CASEI
1TSM 1998-06-17 3.0 L. CASEI THYMIDYLATE SYNTHASE WITH SPECIES SPECIFIC INHIBITOR
1TDA 1993-07-15 3.09 STRUCTURES OF THYMIDYLATE SYNTHASE WITH A C-TERMINAL DELETION: ROLE OF THE C-TERMINUS IN ALIGNMENT OF D/UMP AND CH2H4FOLATE

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Thymidylate synthase P00469 TYSY_LACCA
93.0 Thymidylate synthase Q039F7 TYSY_LACP3