Folding topology and DNA binding of the N-terminal fragment of ada protein

Hitoshi Sakashita, Takahiko Sakuma, Tadayasu Ohkubo, Masatsune Kainosho, Sakumi Kunihiko, Mutsuo Sekiguchi, Kosuke Morikawa

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Three amino terminal fragments of Escherichia coli Ada protein (39 kDa) with different molecular masses (14 kDa, 16 kDa and 20 kDa) were prepared in large quantities from an E. coli strain harboring plasmids constructed for the overproduction of the truncated proteins. The three fragments can be methylated to an extent similar to that of the intact molecule. The methylated 16 kDa fragment specifically binds to the ada box on a DNA duplex. NMR analyses revealed that the 14 kDa fragment comprises two α-helices and a β-sheet with parallel and anti-parallel mixed strands. A comparison of the 15N-1H HMQC spectra of the fragments has led to the conclusion that this tertiary structure within the 14 kDa fragment is retained in the larger 16 kDa and 20 kDa fragments.

Original languageEnglish
Pages (from-to)252-256
Number of pages5
JournalFEBS Letters
Volume323
Issue number3
DOIs
Publication statusPublished - Jun 1 1993

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Escherichia coli Proteins
Escherichia coli
Plasmids
Topology
DNA
Molecular mass
Proteins
Nuclear magnetic resonance
Molecules
vitamin B 12 factor III

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Sakashita, H., Sakuma, T., Ohkubo, T., Kainosho, M., Kunihiko, S., Sekiguchi, M., & Morikawa, K. (1993). Folding topology and DNA binding of the N-terminal fragment of ada protein. FEBS Letters, 323(3), 252-256. https://doi.org/10.1016/0014-5793(93)81351-Y

Folding topology and DNA binding of the N-terminal fragment of ada protein. / Sakashita, Hitoshi; Sakuma, Takahiko; Ohkubo, Tadayasu; Kainosho, Masatsune; Kunihiko, Sakumi; Sekiguchi, Mutsuo; Morikawa, Kosuke.

In: FEBS Letters, Vol. 323, No. 3, 01.06.1993, p. 252-256.

Research output: Contribution to journalArticle

Sakashita, H, Sakuma, T, Ohkubo, T, Kainosho, M, Kunihiko, S, Sekiguchi, M & Morikawa, K 1993, 'Folding topology and DNA binding of the N-terminal fragment of ada protein', FEBS Letters, vol. 323, no. 3, pp. 252-256. https://doi.org/10.1016/0014-5793(93)81351-Y
Sakashita H, Sakuma T, Ohkubo T, Kainosho M, Kunihiko S, Sekiguchi M et al. Folding topology and DNA binding of the N-terminal fragment of ada protein. FEBS Letters. 1993 Jun 1;323(3):252-256. https://doi.org/10.1016/0014-5793(93)81351-Y
Sakashita, Hitoshi ; Sakuma, Takahiko ; Ohkubo, Tadayasu ; Kainosho, Masatsune ; Kunihiko, Sakumi ; Sekiguchi, Mutsuo ; Morikawa, Kosuke. / Folding topology and DNA binding of the N-terminal fragment of ada protein. In: FEBS Letters. 1993 ; Vol. 323, No. 3. pp. 252-256.
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