Altered form of alkaline phosphatase produced by Hep.2 substrain derived from HeLa cells.

HeLa細胞由来のHep. 2細胞株からアルカリ性ホスファターゼ(AP)を, 胎盤型あるいは小腸型APに対する二つのモノクロナール抗体を用いたイムノアフィニティクロマトグラフィーによって精製した. すなわち, 細胞粗抽出液をまず抗小腸AP抗体カラムにのせ, 溶出されたAPを次に抗胎盤APカラムにのせて精製した. 得られたAPはSDS-ポリアクリルアミドゲル電気泳動では二つのサブユニットからなり, 通常のポリアクリルアミドゲル電気泳動では1本の活性バンドが認められた. 17番目までのアミノ酸配列は, Hep. 2細胞APは胎盤型および小腸型APの中間型であった. さらにHep. 2細胞にホスファチジルイノシトール特異的ホスホリパーゼC処理を行うと, 抗胎盤型および抗小腸型AP抗体と反応するAPが等量遊離された. これらの結果から, Hep. 2細胞は胎盤型および小腸型APからなるヘテロダイマーとしてAPを産生していることが考えられた.

[1]  I. Koyama,et al.  Purification and partial characterization of intestinal-like alkaline phosphatase in rabbit kidney. , 1991, Archives of biochemistry and biophysics.

[2]  U. Stendahl,et al.  Purification and characterisation of the placental-like alkaline phosphatase from ovarian epithelial tumours. , 1990, The British journal of cancer. Supplement.

[3]  K. Higashino,et al.  Expression of a hybrid form of alkaline phosphatase isoenzyme in a newly established cell line (HuG-1) from a gastric cancer patient. , 1990, Cancer research.

[4]  K. Higashino,et al.  Structures of the asparagine-linked oligosaccharides of an alkaline phosphatase, kasahara isozyme, purified from FL amnion cells. , 1990, Cancer research.

[5]  U. Stendahl,et al.  Placental-like alkaline phosphatase from ovary and ascites of a patient with a malignant ovarian tumour. , 1990, Clinica chimica acta; international journal of clinical chemistry.

[6]  J. Millán,et al.  Seminoma-derived Nagao isozyme is encoded by a germ-cell alkaline phosphatase gene. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[7]  I. Koyama,et al.  Similarity of the sugar moiety of human alkaline phosphatases between the kidney cortex and duodenum, or medulla and ileum. , 1988, Clinica chimica acta; international journal of clinical chemistry.

[8]  T. Tanaka,et al.  Specific assays for human alkaline phosphatase isozymes. , 1987, Clinica chimica acta; international journal of clinical chemistry.

[9]  S. Udenfriend,et al.  Cloning and sequencing of human intestinal alkaline phosphatase cDNA. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[10]  I. Koyama,et al.  Sugar-chain heterogeneity of human alkaline phosphatases: differences between normal and tumour-associated isozymes. , 1987, Journal of chromatography.

[11]  W. Fishman,et al.  Oncotrophoblast gene expression: placental alkaline phosphatase. , 1987, Advances in cancer research.

[12]  J. Millán.  Molecular cloning and sequence analysis of human placental alkaline phosphatase. , 1986, The Journal of biological chemistry.

[13]  I. Koyama,et al.  Different lectin affinities in rat alkaline phosphatase isozymes: multiple forms of the isozyme isolated by heterogeneities of sugar moieties. , 1986, Journal of chromatography.

[14]  W. Fishman,et al.  Monoclonal antibodies block the bromelain‐mediated release of human placental alkaline phosphatase from cultured cancer cells , 1985, FEBS letters.

[15]  S. Kornfeld,et al.  Assembly of asparagine-linked oligosaccharides. , 1985, Annual review of biochemistry.

[16]  R. Jemmerson,et al.  Monoclonal antibodies block the trypsin cleavage site on human placental alkaline phosphatase , 1984, FEBS letters.

[17]  H. Harris,et al.  Monoclonal antibodies against placental-like and intestinal-like alkaline phosphatases in a malignant human cell line. , 1984, European journal of biochemistry.

[18]  T. Stigbrand.  Present status and future trends of human alkaline phosphatases. , 1984, Progress in clinical and biological research.

[19]  H. Sussman,et al.  Characterization of human foetal intestinal alkaline phosphatase. Comparison with the isoenzymes from the adult intestine and human tumour cell lines. , 1983, The Biochemical journal.

[20]  H. Harris,et al.  Demonstration using monoclonal antibodies of inter-locus heteromeric isozymes of human alkaline phosphatase. , 1982, Journal of immunological methods.

[21]  R. Cummings,et al.  Fractionation of asparagine-linked oligosaccharides by serial lectin-Agarose affinity chromatography. A rapid, sensitive, and specific technique. , 1982, The Journal of biological chemistry.

[22]  H. Harris,et al.  Modulation of the expression of alkaline phosphatase genes in a human malignant cell line passaged through nude mice , 1981, International journal of cancer.

[23]  H. Sussman,et al.  Comparison of human alkaline phosphatase isoenzymes. Structural evidence for three protein classes. , 1979, The Biochemical journal.

[24]  H. Sussman,et al.  Characterization of KB cell alkaline phosphatase. Evidence of similarity to placental alkaline phosphatase. , 1976, The Journal of biological chemistry.

[25]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.