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RISS 인기검색어
- 검색키워드
- 저자 : 김기수 ( Keun Bai Lee (검색결과 2 건)
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종양조직의 Transfer Ribonucleic Acid의 Methyl화에 관한 연구
이근배,이희성,이강석,김기수,Lee, Keun-Bai,Lee, Hi-Sung,Lee, Kang-Suk,Kim, Ki-Sao 생화학분자생물학회 1971 한국생화학회지 Vol.4 No.1
정상조직으로는 동물의 간, 종양조직으로는 Sarcoma 180 ascites tumor cells 및 Walker 256 carcinosarcoma 등을 시료로 사용하여 tRNA를 정제 하였고 또 같은 조직에서 methylase를 추출하여 in vitro에서 tRNA를 homologous 및 heterologous하게 methyl화를 행하여 다음과 같은 몇가지의 결과를 얻었다. 1. Homologous하게 methylate 시켰을 때 종양조직이 정상조직 보다 3 배의 methyl화 능을 나타냈다. 2. 종양조직의 methylase로 정상조직의 tRNA를 기질로 하여 heterologous 하게 methyl화 시켰을 때 정상조직의 homologous methylase에 비하여 4-5배의 methyl 화능을 나타냈다. 특히 Walker 256 carcinosarcoma의 methylase의 활성도가 가장 높았다. 3. 종양조직에 존재하는 tRNA methylase는 guanine 유도체 보다 adenine 유도체를 더 많이 methylate 시켰다. 4. Paper chromatography로 분리 한 methylated bases는 14종이며 그 중 13종의 화학구조가 확인 되었다. 5. 정상조직 및 종양조직에서 문헌에 보고되어 있지 않은 N,N'-dimethylcytosine을 종양조직인 Walker 256 carcinosarcoma에서 분리 확인 하였다. 이 물질은 Walker 256 carcinosarcoma의 methylase로서 다른 종의 tRNA를 methyl화 시켰을 때에도 분리 확인 할 수 있었다. 6. Adenine 유도체로 추정되는 1종류의 methylated bases를 종양조직인 Walker 256 carcinosarcoma 및 Sarcoma 180 ascites tumor cells에서 각각 분리 하였다. Studies on transfer ribonucleic acid (tRNA) methylases of normal tissues, Walker 256 carcinosarcoma and Sarcoma 180 ascites tumor cells have been carried out with respect to: 1) rate of methylation of tRNA from rabbit liver, Walker 256 carcinosarcoma, Sarcoma 180 ascites tumor cells and E. coli Band 2) an attempt was also made to search out the pattern of methylation, i.e., the specific bases methylated by the extracts of tumor tissues. The homogenate was centrifuged at 100,000 xg for 10 min and the supernatant fluid was used as the enzyme extracts. In vitro methylation of tRNA was carried out by incubating enzyme extract with substrate (tRNA) and ATP generating system. $^{14}C$-methyl-L-methionine was used as methyl donor. Resulting methylated tRNA was hydrolyzed and chromatographed on Whatman paper. Spots on the chromatograms were identified by measuring the Rf and UV absorption spectra and cut into strips and counted directly in a Beckman LS-100 liquid scintillation counter. The results obtained were as follows: 1. The tRNA methylases of tumor tissues exhibited the elevaton of methylating potencies about three-fold increase when methylated homologously. 2. The methylase activity of tumor tissues was increased 4-5 times over that of normal controls. The enzyme extracted from Walker 256 carcinosarcoma served as the most appropriate source. 3. Several lines of evidence for the binary nature of the tRNA methylase from tumor tissues were demonstrated. Results of heterologous methylations with a variety of substrate reveal that extracts of tumor tissues introduce more methyl group into adenine residue than into guanine residue of tRNA. 4. Exposure of tRNA to normal methylases followed by methylases from tumor tissues has indicated that the tumor tRNA methylases introduced methyl groups into different positions on the tRNA molecule than do normal enzymes. Among hydrolysate of methylated tRNA, a novel methylated pyrimidine, i.e., N,N'-dimethylcytosine and an unidentified base were detectable. The only extract from Walker 256 carcinosarcoma was found to possess methylase activity for the synthesis of N,N'-dimethylcytosine.
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종양조직의 Transfer Ribonucleic Acid 의 Methyl 화에 관한 연구
이근배,이희성,이강석,김기수 ( Keun Bai Lee,Hi Sung Lee,Kang Suk Lee ) 생화학분자생물학회 1971 BMB Reports Vol.4 No.1
Studies on transfer ribonucleic acid (tRNA) methylases of normal tissues, Walker 256 carcinosarcoma and Sarcoma 180 ascites tumor cells have been carried out with respect to: 1) rate of methylation of tRNA from rabbit liver, Walker 256 carcinosarcoma, Sarcoma 180 ascites tumor cells and E. coli B and 2) an attempt was also made to search out the pattern of methylation, i.e., the specific bases methylated by the extracts of tumor tissues. The homogenate was centrifuged at 100,000 xg for 10 min and the supernatant fluid was used as the enzyme extracts. In vitro methylation of tRNA was carried out by incubating enzyme extract with substrate (tRNA) and ATP generating system. ^(14)C-methyl-L-methionine was used as methyl donor. Resulting methylated tRNA was hydrolyzed and chromatographed on Whatman paper. Spots on the chromatograms were identified by measuring the Rf and UV absorption spectra and cut into strips and counted directly in a Beckman LS-100 liquid scintillation counter. The results obtained were as follows 1. The tRNA methylases of tumor tissues exhibited the elevaton of methylating potencies about three-fold increase when methylated homologously. 2. The methylase activity of tumor tissues was increased 4-5 times over that of normal controls. The enzyme extracted from Walker 256 carcinosarcoma served as the most appropriate source. 3. Several lines of evidence for the binary nature of the tRNA methylase from tumor tissues were demonstrated. Results of heterologous methylations with a variety of substrate reveal that extracts of tumor tissues introduce more methyl group into adenine residue than into guanine residue of tRNA. 4. Exposure of tRNA to normal methylases followed by methylases from tumor tissues has indicated that the tumor tRNA methylases introduced methyl groups into different positions on the tRNA molecule than do normal enzymes. Among hydrolysate of methylated tRNA, a novel methylated pyrimidine, i.e., N,N`-dimethylcytosine and an unidentified base were detectable. The only extract from Walker 256 carcinosarcoma was found to possess methylase activity for the synthesis of N,N`-dimethylcytosine.
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