STIMULATION AND DISORDER OF PHOSPHATIDYLCHOLINE METABOLISM CELL GROWTH
| Project/Area Number |
05454164
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pathological medical chemistry
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| Research Institution | Gunma University |
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Principal Investigator |
YAMASHITA Satoshi Gunma University, Department of Biochemistry, Professor, 医学部, 教授 (50025623)
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| Co-Investigator(Kenkyū-buntansha) |
UCHIDA Tsutomu Gunma University, Department of Biochemistry, Assistant, 医学部, 助手 (00160276)
HOSAKA Kohei Gunma University, Department of Biochemistry, Associate Professor, 医学部, 助教授 (70108992)
SUGIMOTO Hiroyuki Gunma University, Department of Biochemistry, Assistant, 医学部, 助手 (00235897)
OKAMURA Shinichi Gunma University, Department of Biochemisty, Assistant, 医学部, 助手 (20224058)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1994: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1993: ¥5,200,000 (Direct Cost: ¥5,200,000)
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| Keywords | phosphatidylcholine metabolism / cell growth / signal transduction / enzyme purification / phospholipase D / lysophospholipase / phosphatidic acid / lysophosphatidylcholine / リゾホスファチジルコン / コリンキナーゼ |
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| Research Abstract |
Phosphatidylcholine is profoundly involved in the signal transduction and growth of cells. We are interested in the roles of phosphatidylcholine metabolism in cell proliferation. The presen study dealt with the purification, and cloning of cDNA of phospholipases in hosphatidylcholine metabolism. The major results can be summarized as follows.
Phospholipase D,believed to play a central role in the cell growth signaling pathway, was purified 2,200-fold to homogeneity from pig liver microsomes by a combination of column chromatographies. The molecular weight was determined to be 190,000 by SDS electrophoresis. Lysophospholipases acting on lysophosphatidylcholine were purified to homogeneity from rat liver and pig gastric mucosa. From liver, two isoforms with molecular masses of 24 and 60 kd were obtained. The latter enzyme exhibited not only hydrolase activity, but also transacylase activity. From gastric mucosa, two lysophospholipases were also obtained with molecular masses of 22 and 23 kd. Antibodies were raised against hepatic 24-kd and gastric 22- and 23-kd enzymes. Western blot analysis showed that the liver 24-kd enzyme was immunologically related to gastric 22-kd enzyme, but not to liver 60-kd and gastric 23-kd lysophospholipases. All these lysophospholipases were inhibited by phosphatidic acid and lysophosphatidic acid to varying extents. We obtained cDNA clone for hepatic 24-kd lysophospholipase from lambda gtl 1 liver cDNA library using the antibody. The cDNA clone contained the nucleotide sequence corresponding to the partial amino acid sequences determined for the purified hepatic 24-kd lysophospholipase.
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Report
(3 results)
Research Products
(21 results)
