Site-directed mutagenesis of the alpha subunit of tryptophan synthase from Salmonella typhimurium.


Abstract

Site-specific mutagenesis has been used to prepare two mutant forms of the alpha subunit of tryptophan synthase from Salmonella typhimurium in which either cysteine-81 or cysteine-118 is replaced by a serine residue. These mutant proteins are potentially useful for x-ray crystallographic studies since a heavy metal binding site is specifically eliminated in each mutant. The purified mutant proteins are fully active in four reactions catalyzed by the wild type alpha 2 beta 2 complex of tryptophan synthase. However, the mutant alpha 2 beta 2 complexes dissociate more readily and are less heat-stable than the wild type alpha 2 beta 2 complex. Thus, cysteine-81 and cysteine-118 of the alpha subunit serve structural but not functional roles. Study holds ProTherm entries: 102, 103, 104 Extra Details: 2-mercaptoethanol (10 mM) and EDTA(1 mM) are added as additives. alpha subunit of tryptophan synthase; site-specific mutagenesis;,structural role; heavy metal binding site

Submission Details

ID: fVMDGtY23

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:14 p.m.

Version: 1

Publication Details
Ahmed SA;Kawasaki H;Bauerle R;Morita H;Miles EW,Biochem. Biophys. Res. Commun. (1988) Site-directed mutagenesis of the alpha subunit of tryptophan synthase from Salmonella typhimurium. PMID:3126743
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
1A50 1998-02-18T00:00:00+0000 2.3 CRYSTAL STRUCTURE OF WILD-TYPE TRYPTOPHAN SYNTHASE COMPLEXED WITH 5-FLUOROINDOLE PROPANOL PHOSPHATE
1A5A 1998-02-12T00:00:00+0000 1.9 CRYO-CRYSTALLOGRAPHY OF A TRUE SUBSTRATE, INDOLE-3-GLYCEROL PHOSPHATE, BOUND TO A MUTANT (ALPHAD60N) TRYPTOPHAN SYNTHASE ALPHA2BETA2 COMPLEX REVEALS THE CORRECT ORIENTATION OF ACTIVE SITE ALPHA GLU 49
1A5B 1998-02-12T00:00:00+0000 2.0 CRYO-CRYSTALLOGRAPHY OF A TRUE SUBSTRATE, INDOLE-3-GLYCEROL PHOSPHATE, BOUND TO A MUTANT (ALPHA D60N) TRYPTOPHAN SYNTHASE ALPHA2BETA2 COMPLEX REVEALS THE CORRECT ORIENTATION OF ACTIVE SITE ALPHA GLU 49
1A5S 1998-02-17T00:00:00+0000 2.3 CRYSTAL STRUCTURE OF WILD-TYPE TRYPTOPHAN SYNTHASE COMPLEXED WITH 5-FLUOROINDOLE PROPANOL PHOSPHATE AND L-SER BOUND AS AMINO ACRYLATE TO THE BETA SITE
1BEU 1998-05-18T00:00:00+0000 1.9 TRP SYNTHASE (D60N-IPP-SER) WITH K+
1BKS 1998-07-10T00:00:00+0000 2.2 TRYPTOPHAN SYNTHASE (E.C.4.2.1.20) FROM SALMONELLA TYPHIMURIUM
1C29 1999-07-23T00:00:00+0000 2.3 CRYSTAL STRUCTURE OF THE COMPLEX OF BACTERIAL TRYPTOPHAN SYNTHASE WITH THE TRANSITION STATE ANALOGUE INHIBITOR 4-(2-HYDROXYPHENYLTHIO)-1-BUTENYLPHOSPHONIC ACID
1C8V 1999-07-30T00:00:00+0000 2.2 CRYSTAL STRUCTURE OF THE COMPLEX OF BACTERIAL TRYPTOPHAN SYNTHASE WITH THE TRANSITION STATE ANALOGUE INHIBITOR 4-(2-HYDROXYPHENYLTHIO)-BUTYLPHOSPHONIC ACID
1C9D 1999-08-02T00:00:00+0000 2.3 CRYSTAL STRUCTURE OF THE COMPLEX OF BACTERIAL TRYPTOPHAN SYNTHASE WITH THE TRANSITION STATE ANALOGUE INHIBITOR 4-(2-HYDROXY-4-FLUOROPHENYLTHIO)-BUTYLPHOSPHONIC ACID
1CW2 1999-08-25T00:00:00+0000 2.0 CRYSTAL STRUCTURE OF THE COMPLEX OF BACTERIAL TRYPTOPHAN SYNTHASE WITH THE TRANSITION STATE ANALOGUE INHIBITOR 4-(2-HYDROXYPHENYLSULFINYL)-BUTYLPHOSPHONIC ACID

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
95.1 Tryptophan synthase alpha chain A9MPY6 TRPA_SALAR
98.9 Tryptophan synthase alpha chain B4TX39 TRPA_SALSV
99.3 Tryptophan synthase alpha chain B5BIC0 TRPA_SALPK
99.3 Tryptophan synthase alpha chain Q5PNM0 TRPA_SALPA
99.3 Tryptophan synthase alpha chain C0Q3N3 TRPA_SALPC
99.3 Tryptophan synthase alpha chain Q57NT2 TRPA_SALCH
99.6 Tryptophan synthase alpha chain Q8Z7E0 TRPA_SALTI
99.6 Tryptophan synthase alpha chain A9MWR3 TRPA_SALPB
99.6 Tryptophan synthase alpha chain B4T6X2 TRPA_SALNS
99.6 Tryptophan synthase alpha chain B4TJK9 TRPA_SALHS
99.6 Tryptophan synthase alpha chain B5R6M4 TRPA_SALG2
99.6 Tryptophan synthase alpha chain B5R3P3 TRPA_SALEP
99.6 Tryptophan synthase alpha chain B5FU65 TRPA_SALDC
99.6 Tryptophan synthase alpha chain B5F4M3 TRPA_SALA4
100.0 Tryptophan synthase alpha chain P00929 TRPA_SALTY
90.1 B Tryptophan synthase alpha chain B2VKT2 TRPB_ERWT9
90.9 B Tryptophan synthase alpha chain Q6D4U0 TRPB_PECAS
91.1 B Tryptophan synthase alpha chain A8GF82 TRPB_SERP5
91.6 B Tryptophan synthase alpha chain C6DGZ5 TRPB_PECCP
93.2 B Tryptophan synthase alpha chain A7MMG1 TRPB_CROS8
92.7 B Tryptophan synthase alpha chain A4WB02 TRPB_ENT38
93.7 B Tryptophan synthase alpha chain A6T7W6 TRPB_KLEP7
93.7 B Tryptophan synthase alpha chain B5XT02 TRPB_KLEP3
96.5 B Tryptophan synthase alpha chain Q31ZV4 TRPB_SHIBS
96.5 B Tryptophan synthase alpha chain B2U0F2 TRPB_SHIB3
96.5 B Tryptophan synthase alpha chain B7N474 TRPB_ECOLU
96.5 B Tryptophan synthase alpha chain Q1RCA6 TRPB_ECOUT
96.5 B Tryptophan synthase alpha chain A1AAN1 TRPB_ECOK1
96.5 B Tryptophan synthase alpha chain B7ML77 TRPB_ECO45
96.5 B Tryptophan synthase alpha chain P0A880 TRPB_SHIFL
96.5 B Tryptophan synthase alpha chain Q0T5D5 TRPB_SHIF8
96.5 B Tryptophan synthase alpha chain B6I9X5 TRPB_ECOSE
96.5 B Tryptophan synthase alpha chain P0A879 TRPB_ECOLI
96.5 B Tryptophan synthase alpha chain B1ITJ4 TRPB_ECOLC
96.5 B Tryptophan synthase alpha chain A7ZZJ7 TRPB_ECOHS
96.5 B Tryptophan synthase alpha chain B1XBL0 TRPB_ECODH
96.5 B Tryptophan synthase alpha chain B7LY17 TRPB_ECO8A
96.5 B Tryptophan synthase alpha chain B7L493 TRPB_ECO55
96.5 B Tryptophan synthase alpha chain A7ZL79 TRPB_ECO24
96.7 B Tryptophan synthase alpha chain Q32GS9 TRPB_SHIDS
96.7 B Tryptophan synthase alpha chain B1LH31 TRPB_ECOSM
96.7 B Tryptophan synthase alpha chain Q8FHV9 TRPB_ECOL6
96.7 B Tryptophan synthase alpha chain Q0TIA9 TRPB_ECOL5
96.7 B Tryptophan synthase alpha chain B7MUA0 TRPB_ECO81
96.7 B Tryptophan synthase alpha chain B7NVN1 TRPB_ECO7I
96.7 B Tryptophan synthase alpha chain B7UR67 TRPB_ECO27
96.7 B Tryptophan synthase alpha chain Q3Z109 TRPB_SHISS
96.7 B Tryptophan synthase alpha chain B5YZP1 TRPB_ECO5E
96.7 B Tryptophan synthase alpha chain Q8X7B6 TRPB_ECO57
97.0 B Tryptophan synthase alpha chain B7LS19 TRPB_ESCF3
96.7 B Tryptophan synthase alpha chain A8AG61 TRPB_CITK8
98.2 B Tryptophan synthase alpha chain A9MPY7 TRPB_SALAR
99.5 B Tryptophan synthase alpha chain Q57NT3 TRPB_SALCH
99.7 B Tryptophan synthase alpha chain P0A2K1 TRPB_SALTY
99.7 B Tryptophan synthase alpha chain P0A2K2 TRPB_SALTI
99.7 B Tryptophan synthase alpha chain B4TX38 TRPB_SALSV
99.7 B Tryptophan synthase alpha chain B5BIC1 TRPB_SALPK
99.7 B Tryptophan synthase alpha chain C0Q3N4 TRPB_SALPC
99.7 B Tryptophan synthase alpha chain A9MWR4 TRPB_SALPB
99.7 B Tryptophan synthase alpha chain Q5PD17 TRPB_SALPA
99.7 B Tryptophan synthase alpha chain B4T6X1 TRPB_SALNS
99.7 B Tryptophan synthase alpha chain B4TJK8 TRPB_SALHS
99.7 B Tryptophan synthase alpha chain B5R6M5 TRPB_SALG2
99.7 B Tryptophan synthase alpha chain B5R3P4 TRPB_SALEP
99.7 B Tryptophan synthase alpha chain B5FU66 TRPB_SALDC
99.7 B Tryptophan synthase alpha chain B5F4M4 TRPB_SALA4