Structural study of immunoaffinity-purified DNA polymerase α-DNA primase complex from calf thymus

Katsuyuki Tamai, Kiyohide Kojima, Takamasa Hanaichi, Shigeo Masaki, Motoshi Suzuki, Hayato Umekawa, Shonen Yoshida

Research output: Contribution to journalArticle

154 Citations (Scopus)

Abstract

The DNA polymerase α-DNA primase complex was purified over 17 000-fold to near homogeneity from calf thymus using an immunoaffinity column. Sodium dodecyl sulfate gel electrophoresis revealed three polypeptides with molecular weights of 140, 50 and 47 kDa, in a ratio of 1:2:0.25. The complex showed a sedimentation coefficient of 9.7 S, a Stokes radius of 56 Å and a native molecular weight of 250-260 kDa. Taken together, the data suggest that the calf thymus dNA polymerase α-DNA primase complex is essentially a heterotrimer of large (140 kDa) and small (50 kDa) subunits in a ratio of 1:2, with a globular conformation. Electron-microscopic studies of the complex revealed a spherical particle of 120 Å in diameter, in agreement with the physicochemical results. The binding of the complex to DNA was also demonstrated.

Original languageEnglish
Pages (from-to)263-273
Number of pages11
JournalBBA - Gene Structure and Expression
Volume950
Issue number3
DOIs
Publication statusPublished - 07-09-1988

Fingerprint

DNA Primase
Thymus
DNA-Directed DNA Polymerase
Thymus Gland
Molecular Weight
Molecular weight
Electrophoresis
Sedimentation
Sodium Dodecyl Sulfate
Conformations
Gels
Electrons
Peptides
DNA

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biophysics
  • Biochemistry
  • Genetics

Cite this

Tamai, Katsuyuki ; Kojima, Kiyohide ; Hanaichi, Takamasa ; Masaki, Shigeo ; Suzuki, Motoshi ; Umekawa, Hayato ; Yoshida, Shonen. / Structural study of immunoaffinity-purified DNA polymerase α-DNA primase complex from calf thymus. In: BBA - Gene Structure and Expression. 1988 ; Vol. 950, No. 3. pp. 263-273.
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Structural study of immunoaffinity-purified DNA polymerase α-DNA primase complex from calf thymus. / Tamai, Katsuyuki; Kojima, Kiyohide; Hanaichi, Takamasa; Masaki, Shigeo; Suzuki, Motoshi; Umekawa, Hayato; Yoshida, Shonen.

In: BBA - Gene Structure and Expression, Vol. 950, No. 3, 07.09.1988, p. 263-273.

Research output: Contribution to journalArticle

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T1 - Structural study of immunoaffinity-purified DNA polymerase α-DNA primase complex from calf thymus

AU - Tamai, Katsuyuki

AU - Kojima, Kiyohide

AU - Hanaichi, Takamasa

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AU - Suzuki, Motoshi

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AU - Yoshida, Shonen

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