Catalytically impaired TrpA subunit of tryptophan synthase from Chlamydia trachomatis is an allosteric regulator of TrpB

Michalska, Karolina; Wellington, Samantha; Maltseva, Natalia; Jedrzejczak, Robert; Selem‐Mojica, Nelly; Rosas‐Becerra, L. Rodrigo; Barona‐Gómez, Francisco; Hung, Deborah T.; Joachimiak, Andrzej

doi:10.1002/pro.4143

Catalytically impaired TrpA subunit of tryptophan synthase from Chlamydia trachomatis is an allosteric regulator of TrpB

Journal Article · Wed Jun 16 00:00:00 EDT 2021 · Protein Science

DOI:https://doi.org/10.1002/pro.4143· OSTI ID:1797338

Michalska, Karolina ^[1]; Wellington, Samantha ^[2]; Maltseva, Natalia ^[3]; Jedrzejczak, Robert ^[3]; Selem‐Mojica, Nelly ^[4]; Rosas‐Becerra, L. Rodrigo ^[4]; ^[4]; Hung, Deborah T. ^[2]; ^[5]

Center for Structural Genomics of Infectious Diseases University of Chicago Chicago Illinois USA, Structural Biology Center, X‐ray Science Division Argonne National Laboratory Argonne Illinois USA
Department of Genetics, Broad Institute of MIT and Harvard Cambridge Massachusetts USA
Center for Structural Genomics of Infectious Diseases University of Chicago Chicago Illinois USA
Evolution of Metabolic Diversity Laboratory Unidad de Genómica Avanzada (Langebio) Cinvestav Mexico
Center for Structural Genomics of Infectious Diseases University of Chicago Chicago Illinois USA, Structural Biology Center, X‐ray Science Division Argonne National Laboratory Argonne Illinois USA, Department of Biochemistry and Molecular Biology University of Chicago Chicago Illinois USA

Abstract

Intracellular growth and pathogenesis of Chlamydia species is controlled by the availability of tryptophan, yet the complete biosynthetic pathway for l‐ Trp is absent among members of the genus. Some representatives, however, preserve genes encoding tryptophan synthase, TrpAB – a bifunctional enzyme catalyzing the last two steps in l‐ Trp synthesis. TrpA (subunit α) converts indole‐3‐glycerol phosphate into indole and glyceraldehyde‐3‐phosphate (α reaction). The former compound is subsequently used by TrpB (subunit β) to produce l‐ Trp in the presence of l‐ Ser and a pyridoxal 5′‐phosphate cofactor (β reaction). Previous studies have indicated that in Chlamydia , TrpA has lost its catalytic activity yet remains associated with TrpB to support the β reaction. Here, we provide detailed analysis of the TrpAB from C. trachomatis D/UW‐3/CX, confirming that accumulation of mutations in the active site of TrpA renders it enzymatically inactive, despite the conservation of the catalytic residues. We also show that TrpA remains a functional component of the TrpAB complex, increasing the activity of TrpB by four‐fold. The side chain of non‐conserved βArg267 functions as cation effector, potentially rendering the enzyme less susceptible to the solvent ion composition. The observed structural and functional changes detected herein were placed in a broader evolutionary and genomic context, allowing identification of these mutations in relation to their trp gene contexts in which they occur. Moreover, in agreement with the in vitro data, partial relaxation of purifying selection for TrpA, but not for TrpB, was detected, reinforcing a partial loss of TrpA functions during the course of evolution.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: Consejo Nacional de Ciencia y Tecnología in Mexico (CONACyT); National Institutes of Health (NIH); Royal Society United Kingdom; USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1797338

Alternate ID(s):: OSTI ID: 1819341
OSTI ID: 1831424
OSTI ID: 1797342

Journal Information:: Protein Science, Journal Name: Protein Science Journal Issue: 9 Vol. 30; ISSN 0961-8368

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: United Kingdom

Language:: English

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Catalytically impaired TrpA subunit of tryptophan synthase from Chlamydia trachomatis is an allosteric regulator of TrpB

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