Neutron diffraction from a microgravity-grown crystal reveals the active site hydrogens of the internal aldimine form of tryptophan synthase

Drago, Victoria N.; Devos, Juliette M.; Blakeley, Matthew P.; Forsyth, V. Trevor; Parks, Jerry M.; Kovalevsky, Andrey; Mueser, Timothy C.

doi:10.1016/j.xcrp.2024.101827

Title: Neutron diffraction from a microgravity-grown crystal reveals the active site hydrogens of the internal aldimine form of tryptophan synthase

Journal Article · 11 February 2024 · Cell Reports Physical Science

DOI: https://doi.org/10.1016/j.xcrp.2024.101827 · OSTI ID:2317765

Drago, Victoria N. ^[1]; Devos, Juliette M. ^[2]; Blakeley, Matthew P. ^[3]; Forsyth, V. Trevor ^[4];

^[5];

^[6]

Univ. of Toledo, OH (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Inst. Laue-Langevin (ILL), Grenoble (France); European Molecular Biology Lab. (EMBL), Grenoble (France); European Synchrotron Radiation Facility (ESRF), Grenoble (France)
Inst. Laue-Langevin (ILL), Grenoble (France)
Lund Univ. (Sweden)
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Univ. of Toledo, OH (United States)

Pyridoxal 5′-phosphate (PLP), the biologically active form of vitamin B6, is an essential cofactor in many biosynthetic pathways. The emergence of PLP-dependent enzymes as drug targets and biocatalysts, such as tryptophan synthase (TS), has underlined the demand to understand PLP-dependent catalysis and reaction specificity. The ability of neutron diffraction to resolve the positions of hydrogen atoms makes it an ideal technique to understand how the electrostatic environment and selective protonation of PLP regulates PLP-dependent activities. Facilitated by microgravity crystallization of TS with the Toledo Crystallization Box, we report the 2.1 Å joint X-ray/neutron (XN) structure of TS with PLP in the internal aldimine form. Positions of hydrogens were directly determined in both the α- and β-active sites, including PLP cofactor. The joint XN structure thus provides insight into the selective protonation of the internal aldimine and the electrostatic environment of TS necessary to understand the overall catalytic mechanism.

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Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities (SUF)

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 2317765

Journal Information:: Cell Reports Physical Science, Journal Name: Cell Reports Physical Science Journal Issue: 2 Vol. 5; ISSN 2666-3864

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
neutron diffraction
neutron crystallography
macromolecular crystallography
pyridoxal 5’-phosphate
microgravity crystallization
fold-type II
enzyme mechanism

Title: Neutron diffraction from a microgravity-grown crystal reveals the active site hydrogens of the internal aldimine form of tryptophan synthase

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