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
1BO7 1998-08-10T00:00:00+0000 2.4 THYMIDYLATE SYNTHASE R179T MUTANT
1BO8 1998-08-10T00:00:00+0000 2.4 THYMIDYLATE SYNTHASE R178T MUTANT
1BP0 1998-08-11T00:00:00+0000 2.4 THYMIDYLATE SYNTHASE R23I MUTANT
1BP6 1998-08-13T00:00:00+0000 2.4 THYMIDYLATE SYNTHASE R23I, R179T DOUBLE MUTANT
1BPJ 1998-08-11T00:00:00+0000 2.4 THYMIDYLATE SYNTHASE R178T, R179T DOUBLE MUTANT
1JMF 1997-09-13T00:00:00+0000 2.5 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1JMG 1997-09-13T00:00:00+0000 2.2 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1JMH 1997-09-13T00:00:00+0000 2.5 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1JMI 1997-09-13T00:00:00+0000 2.5 CONTRIBUTIONS OF ORIENTATION AND HYDROGEN BONDING TO CATALYSIS IN ASN-229 MUTANTS OF THYMIDYLATE SYNTHASE
1LCA 1995-06-22T00:00:00+0000 2.5 LACTOBACILLUS CASEI THYMIDYLATE SYNTHASE TERNARY COMPLEX WITH DUMP AND CB3717

Relevant UniProtKB Entries

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