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Role of F225 in O-phosphoserine sulfhydrylase from Aeropyrum pernix K1

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Abstract

O-Phosphoserine sulfhydrylase (OPSS) synthesizes cysteine from O-phospho-l-serine (OPS) and sulfide. We have determined the three-dimensional structures of OPSS from hyperthermophilic archaeon Aeropyrum pernix K1 (ApOPSS) in complex with aminoacrylate intermediate (AA) formed from pyridoxal 5′-phosphate with OPS or in complex with cysteine and compared them with that of ApOPSS. We found an orientational change of F225 at the active-site entrance and constructed an F225A mutant to examine its activities and AA stability and clarify the role of F225 in ApOPSS. The OPS and O-acetyl-l-serine (OAS) sulfhydrylase activities of the F225A mutant decreased by 4.2- and 15-fold compared to those of the wild-type (wt) ApOPSS, respectively. The ability of OPS and OAS to form AA also decreased by 12- and 27-fold, respectively. AA was less stable in the F225A mutant than in the wt ApOPSS. Simulated docking showed that leaving groups, such as phosphate and acetate, were oriented to the inside of the active site in the F225A mutant, whereas they were oriented to the entrance in the wt ApOPSS. These results suggest that F225 in ApOPSS plays important roles in maintaining the hydrophobic environment of AA from solvent water and in controlling the orientation of leaving groups.

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Abbreviations

AA:

Aminoacrylate intermediate

ApOPSS:

OPSS from hyperthermophilic archaeon Aeropyrum pernix K1

OAS:

O-Acetyl-l-serine

OASS:

OAS sulfhydrylase

OPS:

O-Phospho-l-serine

OPSS:

OPS sulfhydrylase

wt:

Wild-type

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Acknowledgments

We are grateful to Drs. Masafumi Shionyu, Tsuyoshi Shirai, Yasushi Kawai, and Yoshisuke Nishi at the Nagahama Institute of Bio-Science and Technology for helpful discussions related to this work. The X-ray diffraction data were obtained on beamline BL44XU (Proposal No. 2009A6928) of SPring-8, Hyogo, Japan with the approval of the Japan Synchrotron Radiation Research Institute. M.K. was supported by the National Institute of Advanced Industrial Science and the Technology Innovation School.

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Authors and Affiliations

  1. Laboratory of Molecular Biochemistry, Nagahama Institute of Bio-Science and Technology, 1266 Tamura-cho, Nagahama, Shiga, 526-0829, Japan

    Emi Takeda, Kohei Kunimoto, Yoshito Kawai & Takashi Nakamura

  2. Biomass Refinery Research Center, National Institute of Advanced Industrial Science and Technology (AIST), 3-11-32 Kagamiyama, Higashihiroshima, Hiroshima, 739-0046, Japan

    Misumi Kataoka & Kazuhiko Ishikawa

  3. Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka, 563-8577, Japan

    Kazuhiko Ishikawa

Authors
  1. Emi Takeda
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  2. Kohei Kunimoto
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  3. Yoshito Kawai
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  4. Misumi Kataoka
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  5. Kazuhiko Ishikawa
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  6. Takashi Nakamura
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Correspondence to Takashi Nakamura.

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Communicated by L. Huang.

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Takeda, E., Kunimoto, K., Kawai, Y. et al. Role of F225 in O-phosphoserine sulfhydrylase from Aeropyrum pernix K1. Extremophiles 20, 733–745 (2016). https://doi.org/10.1007/s00792-016-0862-6

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