A thermodynamic analysis of conformational change due to the alpha 2 beta 2 complex formation of tryptophan synthase.

Abstract

A characteristic property of the tryptophan synthase alpha 2 beta 2 complex is the mutual activation of the alpha and beta subunit upon complex formation. It has been speculated that this mutual activation results from the conformational change due to the alpha/beta subunit interaction. To elucidate this mechanism, we investigated the thermodynamic parameters of association for the various combinations of the alpha and beta subunits from Escherichia coli and Salmonella typhimurium using isothermal titration calorimetry. The negative association enthalpy of the S. typhimurium alpha subunit with the beta subunit from E. coli (or S. typhimurium) was about 20 kJ mol-1 larger than that of the E. coli alpha subunit at 40 degrees C. However, the favorable enthalpy of the S. typhimurium alpha subunit was perfectly compensated by the unfavorable association entropy, therefore, the Gibbs energy of association was similar to that of the E. coli alpha subunit. Furthermore, the site-directed mutagenesis study revealed that a single mutation (K109N; [Asn109] alpha subunit) of the E coli alpha subunit at the subunit interface from E. coli to the S. typhimurium type could change the characteristics of the thermodynamic parameters of association to the S. typhimurium alpha subunit type. The heat-capacity changes of the association of the alpha subunit with the beta subunit were quite great, 6.37-8.21 kJ mol-1 K-1, compared with that due to a decrease in accessible surface area in the subunit interface. The analysis of the thermodynamic parameters of association suggested that the complex formation couples with the folding (rearrangements) of the alpha subunit monomer or/and beta subunit dimer. Study holds ProTherm entries: 7547, 7548, 7549 Extra Details: additive : EDTA(1 mM), calorimetry; thermodynamics; activation;,protein/protein interaction; tryptophan synthase

Submission Details

ID: yfN2WqWQ4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:34 p.m.

Version: 1

Publication Details
Hiraga K;Yutani K,Eur. J. Biochem. (1996) A thermodynamic analysis of conformational change due to the alpha 2 beta 2 complex formation of tryptophan synthase. PMID:8797836
Additional Information

Study Summary

Number of data points 4
Proteins Tryptophan synthase alpha chain ; Tryptophan synthase alpha chain ; Tryptophan synthase alpha chain ; Tryptophan synthase alpha chain
Unique complexes 3
Assays/Quantities/Protocols Experimental Assay: Tm ; Derived Quantity: dTm
Libraries Mutations for sequence MERYESLFAQLKERKEGAFVPFVTLGDPGIEQSLKIIDTLIEAGADALELGIPFSDPLADGPTIQNATLRAFAAGVTPAQCFEMLALIRQKHPTIPIGLLMYANLVFNKGIDEFYAQCEKVGVDSVLVADVPVEESAPFRQAALRHNVAPIFICPPNADDDLLRQIASYGRGYTYLLSRAGVTGAENRAALPLNHLVAKLKEYNAAPPLQGFGISAPDQVKAAIDAGAAGAISGSAIVKIIEQHINEPEKMLAALKVFVQPMKAATRS ; Mutations for sequence A:MERYENLFAQLNDRREGAFVPFVTLGDPGIEQSLKIIDTLIDAGADALELGVPFSDPLADGPTIQNANLRAFAAGVTPAQCFEMLALIREKHPTIPIGLLMYANLVFNNGIDAFYARCEQVGVDSVLVADVPVEESAPFRQAALRHNIAPIFICPPNADDDLLRQVASYGRGYTYLLSRSGVTGAENRGALPLHHLIEKLKEYHAAPALQGFGISSPEQVSAAVRAGAAGAISGSAIVKIIEKNLASPKQMLAELRSFVSAMKAASRA/B:TTLLNPYFGEFGGMYVPQILMPALNQLEEAFVRAQKDPEFQAQFADLLKNYAGRPTALTKCQNITAGTRTTLYLKREDLLHGGAHKTNQVLGQALLAKRMGKSEIIAETGAGQHGVASALASALLGLKCRIYMGAKDVERQSPNVFRMRLMGAEVIPVHSGSATLKDACNEALRDWSGSYETAHYMLGTAAGPHPYPTIVREFQRMIGEETKAQILDKEGRLPDAVIACVGGGSNAIGMFADFINDTSVGLIGVEPGGHGIETGEHGAPLKHGRVGIYFGMKAPMMQTADGQIEESYSISAGLDFPSVGPQHAYLNSIGRADYVSITDDEALEAFKTLCRHEGIIPALESSHALAHALKMMREQPEKEQLLVVNLSGRGDKDIFTVHDILKARGEI

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 UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Tryptophan synthase alpha chain P00929 TRPA_SALTY
99.6 Tryptophan synthase alpha chain B5F4M3 TRPA_SALA4
99.6 Tryptophan synthase alpha chain B5FU65 TRPA_SALDC
99.6 Tryptophan synthase alpha chain B5R3P3 TRPA_SALEP
99.6 Tryptophan synthase alpha chain B5R6M4 TRPA_SALG2
99.6 Tryptophan synthase alpha chain B4TJK9 TRPA_SALHS
99.6 Tryptophan synthase alpha chain B4T6X2 TRPA_SALNS
99.6 Tryptophan synthase alpha chain A9MWR3 TRPA_SALPB
99.6 Tryptophan synthase alpha chain Q8Z7E0 TRPA_SALTI
99.3 Tryptophan synthase alpha chain Q57NT2 TRPA_SALCH
99.3 Tryptophan synthase alpha chain C0Q3N3 TRPA_SALPC
99.3 Tryptophan synthase alpha chain Q5PNM0 TRPA_SALPA
99.3 Tryptophan synthase alpha chain B5BIC0 TRPA_SALPK
98.9 Tryptophan synthase alpha chain B4TX39 TRPA_SALSV
95.1 Tryptophan synthase alpha chain A9MPY6 TRPA_SALAR
99.7 B Tryptophan synthase alpha chain B5F4M4 TRPB_SALA4
99.7 B Tryptophan synthase alpha chain B5FU66 TRPB_SALDC
99.7 B Tryptophan synthase alpha chain B5R3P4 TRPB_SALEP
99.7 B Tryptophan synthase alpha chain B5R6M5 TRPB_SALG2
99.7 B Tryptophan synthase alpha chain B4TJK8 TRPB_SALHS
99.7 B Tryptophan synthase alpha chain B4T6X1 TRPB_SALNS
99.7 B Tryptophan synthase alpha chain Q5PD17 TRPB_SALPA
99.7 B Tryptophan synthase alpha chain A9MWR4 TRPB_SALPB
99.7 B Tryptophan synthase alpha chain C0Q3N4 TRPB_SALPC
99.7 B Tryptophan synthase alpha chain B5BIC1 TRPB_SALPK
99.7 B Tryptophan synthase alpha chain B4TX38 TRPB_SALSV
99.7 B Tryptophan synthase alpha chain P0A2K2 TRPB_SALTI
99.7 B Tryptophan synthase alpha chain P0A2K1 TRPB_SALTY
99.5 B Tryptophan synthase alpha chain Q57NT3 TRPB_SALCH
98.2 B Tryptophan synthase alpha chain A9MPY7 TRPB_SALAR
96.7 B Tryptophan synthase alpha chain A8AG61 TRPB_CITK8
97.0 B Tryptophan synthase alpha chain B7LS19 TRPB_ESCF3
96.7 B Tryptophan synthase alpha chain Q8X7B6 TRPB_ECO57
96.7 B Tryptophan synthase alpha chain B5YZP1 TRPB_ECO5E
96.7 B Tryptophan synthase alpha chain Q3Z109 TRPB_SHISS
96.7 B Tryptophan synthase alpha chain B7UR67 TRPB_ECO27
96.7 B Tryptophan synthase alpha chain B7NVN1 TRPB_ECO7I
96.7 B Tryptophan synthase alpha chain B7MUA0 TRPB_ECO81
96.7 B Tryptophan synthase alpha chain Q0TIA9 TRPB_ECOL5
96.7 B Tryptophan synthase alpha chain Q8FHV9 TRPB_ECOL6
96.7 B Tryptophan synthase alpha chain B1LH31 TRPB_ECOSM
96.7 B Tryptophan synthase alpha chain Q32GS9 TRPB_SHIDS
96.5 B Tryptophan synthase alpha chain A7ZL79 TRPB_ECO24
96.5 B Tryptophan synthase alpha chain B7L493 TRPB_ECO55
96.5 B Tryptophan synthase alpha chain B7LY17 TRPB_ECO8A
96.5 B Tryptophan synthase alpha chain B1XBL0 TRPB_ECODH
96.5 B Tryptophan synthase alpha chain A7ZZJ7 TRPB_ECOHS
96.5 B Tryptophan synthase alpha chain B1ITJ4 TRPB_ECOLC
96.5 B Tryptophan synthase alpha chain P0A879 TRPB_ECOLI
96.5 B Tryptophan synthase alpha chain B6I9X5 TRPB_ECOSE
96.5 B Tryptophan synthase alpha chain Q0T5D5 TRPB_SHIF8
96.5 B Tryptophan synthase alpha chain P0A880 TRPB_SHIFL
96.5 B Tryptophan synthase alpha chain B7ML77 TRPB_ECO45
96.5 B Tryptophan synthase alpha chain A1AAN1 TRPB_ECOK1
96.5 B Tryptophan synthase alpha chain Q1RCA6 TRPB_ECOUT
96.5 B Tryptophan synthase alpha chain B7N474 TRPB_ECOLU
96.5 B Tryptophan synthase alpha chain B2U0F2 TRPB_SHIB3
96.5 B Tryptophan synthase alpha chain Q31ZV4 TRPB_SHIBS
93.7 B Tryptophan synthase alpha chain B5XT02 TRPB_KLEP3
93.7 B Tryptophan synthase alpha chain A6T7W6 TRPB_KLEP7
92.7 B Tryptophan synthase alpha chain A4WB02 TRPB_ENT38
93.2 B Tryptophan synthase alpha chain A7MMG1 TRPB_CROS8
91.6 B Tryptophan synthase alpha chain C6DGZ5 TRPB_PECCP
91.1 B Tryptophan synthase alpha chain A8GF82 TRPB_SERP5
90.9 B Tryptophan synthase alpha chain Q6D4U0 TRPB_PECAS
90.1 B Tryptophan synthase alpha chain B2VKT2 TRPB_ERWT9
100.0 Tryptophan synthase alpha chain C4ZTV3 TRPA_ECOBW
100.0 Tryptophan synthase alpha chain B1XBK9 TRPA_ECODH
100.0 Tryptophan synthase alpha chain A7ZZJ6 TRPA_ECOHS
100.0 Tryptophan synthase alpha chain B1ITJ5 TRPA_ECOLC
100.0 Tryptophan synthase alpha chain P0A877 TRPA_ECOLI
100.0 Tryptophan synthase alpha chain P0A878 TRPA_SHIFL
99.6 Tryptophan synthase alpha chain B7LY16 TRPA_ECO8A
99.6 Tryptophan synthase alpha chain B6I9X4 TRPA_ECOSE
99.6 Tryptophan synthase alpha chain B7L492 TRPA_ECO55
99.3 Tryptophan synthase alpha chain A7ZL78 TRPA_ECO24
99.6 Tryptophan synthase alpha chain B2U0F1 TRPA_SHIB3
99.3 Tryptophan synthase alpha chain Q8FHW0 TRPA_ECOL6
99.6 Tryptophan synthase alpha chain Q31ZV3 TRPA_SHIBS
99.6 Tryptophan synthase alpha chain Q0T5D6 TRPA_SHIF8
98.9 Tryptophan synthase alpha chain Q3Z108 TRPA_SHISS
99.3 Tryptophan synthase alpha chain Q32GT0 TRPA_SHIDS
98.9 Tryptophan synthase alpha chain Q8X7B5 TRPA_ECO57
98.9 Tryptophan synthase alpha chain B5YZP0 TRPA_ECO5E
98.5 Tryptophan synthase alpha chain B7ML76 TRPA_ECO45
98.5 Tryptophan synthase alpha chain A1AAN0 TRPA_ECOK1
98.5 Tryptophan synthase alpha chain Q1RCA7 TRPA_ECOUT
98.9 Tryptophan synthase alpha chain B7UR66 TRPA_ECO27
98.1 Tryptophan synthase alpha chain B7MU99 TRPA_ECO81
98.5 Tryptophan synthase alpha chain Q0TIB0 TRPA_ECOL5
98.5 Tryptophan synthase alpha chain B7N473 TRPA_ECOLU
98.1 Tryptophan synthase alpha chain B1LH32 TRPA_ECOSM
97.8 Tryptophan synthase alpha chain B7NVN0 TRPA_ECO7I
97.0 Tryptophan synthase alpha chain B7LS20 TRPA_ESCF3