Purine but not Pyrimidine Nucleotides Support Rotation of F 1-ATPase

Hiroyuki Noji, Dirk Bald, Ryohei Yasuda, Hiroyasu Itoh, Masasuke Yoshida, Kazuhiko Kinosita

研究成果: Article

46 引用 (Scopus)

抄録

The binding change model for the F1,-ATPase predicts that its rotation is intimately correlated with the changes in the affinities of the three catalytic sites for nucleotides. If so, subtle differences in the nucleotide structure may have pronounced effects on rotation. Here we show by single-molecule imaging that purine nucleotides ATP, GTP, and ITP support rotation but pyrimidine nucleotides UTP and CTP do not, suggesting that the extra ring in purine is indispensable for proper operation of this molecular motor. Although the three purine nucleotides were bound to the enzyme at different rates, all showed similar rotational characteristics: counterclockwise rotation, 120° steps each driven by hydrolysis of one nucleotide molecule, occasional back steps, rotary torque of ∼40 piconewtons (pN)·nm, and mechanical work done in a step of ∼80 pN·nm. These latter characteristics are likely to be determined by the rotational mechanism built in the protein structure, which purine nucleotides can energize. With ATP and GTP, rotation was observed even when the free energy of hydrolysis was-80 pN·nm/molecule, indicating -100% efficiency. Reconstituted FoF1-ATPase actively translocated protons by hydrolyzing ATP, GTP, and ITP, but CTP and UTP were not even hydrolyzed. Isolated F1 very slowly hydrolyzed UTP (but not CTP), suggesting possible uncoupling from rotation.

元の言語English
ページ(範囲)25480-25486
ページ数7
ジャーナルJournal of Biological Chemistry
276
発行部数27
DOI
出版物ステータスPublished - 2001 7 6
外部発表Yes

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Pyrimidine Nucleotides
Proton-Translocating ATPases
Purine Nucleotides
Cytidine Triphosphate
Uridine Triphosphate
Guanosine Triphosphate
Inosine Triphosphate
Nucleotides
Adenosine Triphosphate
Molecules
Hydrolysis
Torque
Free energy
Adenosine Triphosphatases
purine
Protons
Catalytic Domain
Imaging techniques
Enzymes

ASJC Scopus subject areas

  • Biochemistry

これを引用

Noji, H., Bald, D., Yasuda, R., Itoh, H., Yoshida, M., & Kinosita, K. (2001). Purine but not Pyrimidine Nucleotides Support Rotation of F 1-ATPase. Journal of Biological Chemistry, 276(27), 25480-25486. https://doi.org/10.1074/jbc.M102200200

Purine but not Pyrimidine Nucleotides Support Rotation of F 1-ATPase. / Noji, Hiroyuki; Bald, Dirk; Yasuda, Ryohei; Itoh, Hiroyasu; Yoshida, Masasuke; Kinosita, Kazuhiko.

:: Journal of Biological Chemistry, 巻 276, 番号 27, 06.07.2001, p. 25480-25486.

研究成果: Article

Noji, H, Bald, D, Yasuda, R, Itoh, H, Yoshida, M & Kinosita, K 2001, 'Purine but not Pyrimidine Nucleotides Support Rotation of F 1-ATPase', Journal of Biological Chemistry, 巻. 276, 番号 27, pp. 25480-25486. https://doi.org/10.1074/jbc.M102200200
Noji, Hiroyuki ; Bald, Dirk ; Yasuda, Ryohei ; Itoh, Hiroyasu ; Yoshida, Masasuke ; Kinosita, Kazuhiko. / Purine but not Pyrimidine Nucleotides Support Rotation of F 1-ATPase. :: Journal of Biological Chemistry. 2001 ; 巻 276, 番号 27. pp. 25480-25486.
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