Identification of the ferredoxin interaction sites on ferredoxin-dependent glutamate synthase from Synechocystis sp. PCC 6803

Masakazu Hirasawa, Jacaranda Solis, Nanditha Vaidyanathan, Anurag P. Srivastava, R. Max Wynn, Roger B. Sutton, David B. Knaff

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Based on in silico docking methods, five amino acids in glutamate synthase (Gln-467, His-1144, Asn-1147, Arg-1162, and Trp-676) likely constitute key binding residues in the interface of a glutamate synthase:ferredoxin complex. Although all interfacial mutants studied showed the ability to form a complex under low ionic strength, these docking mutations showed significantly less ferredoxin-dependent activities, while still retaining enzymatic activity. Furthermore, isothermal titration calorimetry showed a possible 1:2 molar ratio between the wild-type glutamate synthase and ferredoxin. However, each of our interfacial mutants showed only a 1:1 complex with ferredoxin, suggesting that the mutations directly affect the glutamate synthase:ferredoxin heterodimer interface.

Original languageEnglish (US)
Pages (from-to)1-12
Number of pages12
JournalPhotosynthesis Research
DOIs
StateAccepted/In press - Oct 3 2017

Fingerprint

glutamate synthase (ferredoxin)
Synechocystis sp. PCC 6803
Synechocystis
Ferredoxins
ferredoxins
Glutamate Synthase
Calorimetry
Mutation
Ionic strength
Titration
Computer Simulation
Osmolar Concentration
mutation
mutants
calorimetry
Amino Acids
ionic strength
titration
amino acids

Keywords

  • Electrostatic interactions
  • Ferredoxin
  • Flavin mononucleotide (FMN)
  • Glutamate synthase
  • In silico docking
  • Iron–sulfur-binding sites
  • Isothermal titration calorimetry
  • Site-directed mutagenesis
  • Spectral perturbation

ASJC Scopus subject areas

  • Biochemistry
  • Plant Science
  • Cell Biology

Cite this

Identification of the ferredoxin interaction sites on ferredoxin-dependent glutamate synthase from Synechocystis sp. PCC 6803. / Hirasawa, Masakazu; Solis, Jacaranda; Vaidyanathan, Nanditha; Srivastava, Anurag P.; Wynn, R. Max; Sutton, Roger B.; Knaff, David B.

In: Photosynthesis Research, 03.10.2017, p. 1-12.

Research output: Contribution to journalArticle

Hirasawa, Masakazu ; Solis, Jacaranda ; Vaidyanathan, Nanditha ; Srivastava, Anurag P. ; Wynn, R. Max ; Sutton, Roger B. ; Knaff, David B. / Identification of the ferredoxin interaction sites on ferredoxin-dependent glutamate synthase from Synechocystis sp. PCC 6803. In: Photosynthesis Research. 2017 ; pp. 1-12.
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abstract = "Based on in silico docking methods, five amino acids in glutamate synthase (Gln-467, His-1144, Asn-1147, Arg-1162, and Trp-676) likely constitute key binding residues in the interface of a glutamate synthase:ferredoxin complex. Although all interfacial mutants studied showed the ability to form a complex under low ionic strength, these docking mutations showed significantly less ferredoxin-dependent activities, while still retaining enzymatic activity. Furthermore, isothermal titration calorimetry showed a possible 1:2 molar ratio between the wild-type glutamate synthase and ferredoxin. However, each of our interfacial mutants showed only a 1:1 complex with ferredoxin, suggesting that the mutations directly affect the glutamate synthase:ferredoxin heterodimer interface.",
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AU - Hirasawa, Masakazu

AU - Solis, Jacaranda

AU - Vaidyanathan, Nanditha

AU - Srivastava, Anurag P.

AU - Wynn, R. Max

AU - Sutton, Roger B.

AU - Knaff, David B.

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