Thermal stability of dihydrofolate reductase and its fused proteins with oligopeptides.


Abstract

Two fused proteins of dihydrofolate reductase (DHFR) with oligopeptides were prepared by a recombinant DNA method. One of these, DHFR-IQI, has three (Ile-Gln-Ile) and the other, DHFR-lek, has eight (Ile-Arg-Met-Tyr-Gly-Gly-Phe-Leu) additional amino acid residues at the C terminals; in both proteins, Cys152 of wild DHFR is replaced by Glu. The thermal transition of the proteins was measured by CD and DSC at pH 7.0 and compared with that of wild DHFR. The results show that the thermal stability of DHFR-IQI is the same as that of the wild DHFR and that of DHFR-lek is less than that of the former two DHFRs. Analysis of the DSC data of DHFR-IQI indicates that the thermal transition is a three-state one. Data from both DSC and CD measurements suggest the association of DHFR-lek molecules. Study holds ProTherm entries: 2803, 2804 Extra Details: additive : EDTA(0.2 mM),CD measurements are at 220nm Escherichia coli dihydrofolate reductase; Thermal stability

Submission Details

ID: cvoYbBZC4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:20 p.m.

Version: 1

Publication Details
Uedaira H;Kidokoro S;Iwakura M;Honda S;Ohashi S,Ann. N. Y. Acad. Sci. (1990) Thermal stability of dihydrofolate reductase and its fused proteins with oligopeptides. PMID:2075977
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
6CYV 2018-04-06T00:00:00+0000 1.3 E. coli DHFR ternary complex with NADP and dihydrofolate
6CW7 2018-03-30T00:00:00+0000 1.03 E. coli DHFR product complex with (6S)-5,6,7,8-TETRAHYDROFOLATE
6CXK 2018-04-03T00:00:00+0000 1.11 E. coli DHFR substrate complex with Dihydrofolate
5UII 2017-01-14T00:00:00+0000 1.35 structure of DHFR with bound buformin and NADP
5UIH 2017-01-14T00:00:00+0000 1.65 structure of DHFR with bound phenformin and NADP
5CC9 2015-07-01T00:00:00+0000 1.2 L28F E.coli dihydrofolate reductase complexed with 5,10-dideazatetrahydrofolate and oxidized nicotinamide adenine dinucleotide phosphate
1RC4 1996-11-25T00:00:00+0000 1.9 DIHYDROFOLATE REDUCTASE COMPLEXED WITH 5,10-DIDEAZATETRAHYDROFOLATE AND NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE (OXIDIZED FORM)
6MR9 2018-10-12T00:00:00+0000 1.35 E. coli DHFR complex with a reaction intermediate
7D3Z 2020-09-21T00:00:00+0000 1.65 X-ray crystal Structure of E.coli Dihydrofolate Reductase complexed with folate and NADP+ at pH4.5
1RX6 1996-09-19T00:00:00+0000 2.0 DIHYDROFOLATE REDUCTASE (E.C.1.5.1.3) COMPLEXED WITH 5,10-DIDEAZATETRAHYDROFOLATE AND NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE (REDUCED FORM)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
91.8 Dihydrofolate reductase P31074 DYR_KLEAE
96.2 Dihydrofolate reductase P31073 DYR_CITFR
100.0 Dihydrofolate reductase P0ABQ6 DYR_SHIFL
100.0 Dihydrofolate reductase P0ABQ4 DYR_ECOLI
100.0 Dihydrofolate reductase P0ABQ5 DYR_ECOL6