TY - JOUR
T1 - Structural study of immunoaffinity-purified DNA polymerase α-DNA primase complex from calf thymus
AU - Tamai, Katsuyuki
AU - Kojima, Kiyohide
AU - Hanaichi, Takamasa
AU - Masaki, Shigeo
AU - Suzuki, Motoshi
AU - Umekawa, Hayato
AU - Yoshida, Shonen
N1 - Funding Information:
The authors thank Mr. Y. Yamazaki of Aichi Prefectural Colony for the preliminary work of the electron microscopic study and Mrs. R. Morishita-Suzuki for purifing calf thymus 10 S DNA polymerase a. This work was supported in part by Grant-in-aid for Cancer Research from the Ministry of Education, Science and Culture of Japan. A part of this study was performed in the Central Laboratory of Radioisotopes in Nagoya University School of Medicine.
PY - 1988/9/7
Y1 - 1988/9/7
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/0023719578
UR - https://www.scopus.com/pages/publications/0023719578#tab=citedBy
U2 - 10.1016/0167-4781(88)90122-4
DO - 10.1016/0167-4781(88)90122-4
M3 - Article
C2 - 3167053
AN - SCOPUS:0023719578
SN - 0167-4781
VL - 950
SP - 263
EP - 273
JO - BBA - Gene Structure and Expression
JF - BBA - Gene Structure and Expression
IS - 3
ER -