Role of the N-terminal extension of the (betaalpha)8-barrel enzyme indole-3-glycerol phosphate synthase for its fold, stability, and catalytic activity.


Abstract

Indole-3-glycerol phosphate synthase (IGPS) catalyzes the fifth step in the biosynthesis of tryptophan. It belongs to the large and versatile family of (betaalpha)(8)-barrel enzymes but has an unusual N-terminal extension of about 40 residues. Limited proteolysis with trypsin of IGPS from both Sulfolobus solfataricus (sIGPS) and Thermotoga maritima (tIGPS) removes about 25 N-terminal residues and one of the two extra helices contained therein. To assess the role of the extension, the N-terminally truncated variants sIGPSDelta(1-26) and tIGPSDelta(1-25) were produced recombinantly in Escherichia coli, purified, and characterized in comparison to the wild-type enzymes. Both sIGPSDelta(1-26) and tIGPSDelta(1-25) have unchanged oligomerization states and turnover numbers. In contrast, their Michaelis constants for the substrate 1-(o-carboxyphenylamino)-1-deoxyribulose 5-phosphate are increased, and their resistance toward unfolding induced by heat and guanidinium chloride is decreased. sIGPSDelta(1-26) was crystallized, and its X-ray structure was solved at 2.8 A resolution. The comparison with the known structure of sIGPS reveals small differences that account for its reduced substrate affinity and protein stability. The structure of the core of sIGPSDelta(1-26) is, however, unchanged compared to sIGPS, explaining its retained catalytic activity and consistent with the idea that it evolved from the same ancestor as the phosphoribosyl anthranilate isomerase and the alpha-subunit of tryptophan synthase. These (betaalpha)(8)-barrel enzymes catalyze the reactions preceding and following IGPS in tryptophan biosynthesis but lack an N-terminal extension. Study holds ProTherm entries: 18881, 18882, 18883, 18884 Extra Details: (betaalpha)(8)-barrel enzymes; substrate affinity; protein stability; tryptophan biosynthesis

Submission Details

ID: gq6ZqVy63

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:50 p.m.

Version: 1

Publication Details
Schneider B;Knöchel T;Darimont B;Hennig M;Dietrich S;Babinger K;Kirschner K;Sterner R,Biochemistry (2005) Role of the N-terminal extension of the (betaalpha)8-barrel enzyme indole-3-glycerol phosphate synthase for its fold, stability, and catalytic activity. PMID:16342933
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
1I4N 2001-02-22T00:00:00+0000 2.5 CRYSTAL STRUCTURE OF INDOLEGLYCEROL PHOSPHATE SYNTHASE FROM THERMOTOGA MARITIMA
1J5T 2002-07-03T00:00:00+0000 3.0 Crystal structure of indole-3-glycerol phosphate synthase (TM0140) from Thermotoga maritima at 3.0 A resolution
1A53 1998-02-19T00:00:00+0000 2.0 COMPLEX OF INDOLE-3-GLYCEROLPHOSPHATE SYNTHASE FROM SULFOLOBUS SOLFATARICUS WITH INDOLE-3-GLYCEROLPHOSPHATE AT 2.0 A RESOLUTION
1IGS 1995-08-11T00:00:00+0000 2.0 INDOLE-3-GLYCEROLPHOSPHATE SYNTHASE FROM SULFOLOBUS SOLFATARICUS AT 2.0 A RESOLUTION
1JUK 1996-05-03T00:00:00+0000 2.5 INDOLE-3-GLYCEROLPHOSPHATE SYNTHASE FROM SULFOLOBUS SOLFATARICUS IN A TRIGONAL CRYSTAL FORM
1JUL 1996-05-03T00:00:00+0000 2.0 INDOLE-3-GLYCEROLPHOSPHATE SYNTHASE FROM SULFOLOBUS SOLFATARICUS IN A SECOND ORTHORHOMBIC CRYSTAL FORM
1LBF 2002-04-03T00:00:00+0000 2.05 CRYSTAL STRUCTURE OF INDOLE-3-GLYCEROL PHOSPHATE SYNTASE (IGPS)WITH REDUCED 1-(O-CABOXYPHENYLAMINO)-1-DEOXYRIBULOSE 5-PHOSPHATE (RCDRP)
1LBL 2002-04-04T00:00:00+0000 2.4 Crystal structure of indole-3-glycerol phosphate synthase (IGPS) in complex with 1-(o-carboxyphenylamino)-1-deoxyribulose 5'-phosphate (CdRP)
2C3Z 2005-10-13T00:00:00+0000 2.8 Crystal structure of a truncated variant of indole-3-glycerol phosphate synthase from Sulfolobus solfataricus
3NYZ 2010-07-15T00:00:00+0000 1.51 Crystal Structure of Kemp Elimination Catalyst 1A53-2

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Indole-3-glycerol phosphate synthase Q56319 TRPC_THEMA
100.0 Indole-3-glycerol phosphate synthase Q06121 TRPC_SACS2
100.0 Multifunctional tryptophan biosynthesis protein P25170 TRPG_PHACH