생명과학회지 (Journal of Life Science)

  • 제8권1호
  • /
  • Pages.14-26
  • /
  • 1998
  • /
  • 1225-9918(pISSN)
  • /
  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지

쥐 소뇌에 있어서 raf protein kinases 의 면역세포 화학적 분포와 단백질 양상

Immunocytochemical distribution of raf protein kinases and protein pattern in rat cerebellum

  • 박정순 (부산대학교 자연과학대학 생물학과) ;
  • 최원철 (부산대학교 자연과학대학 생물학과)
  • 발행 : 1998.02.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

본 연구에서는 SDS/polyacrylamide 젤 전기영동에 의한 쥐의 성장과정에 따른 소뇌의 단백질양상의 변화양상과 immunocytochemistry를 이용하여 c-raf a-raf kinase의 정상 소뇌에서의 분포에 대해 관찰 하였으며 western blot을 이용하여 소뇌의 단백질들에서 c-raf의 존재에 대해 살펴보았다. 단백질 양상에서 쥐의 성장에 따라 crude에선,ㄴ 49,200 dalton과 169,000 dalton 사이의 bands가 양적 증가를 보였으며 cytosolic fraction 에서는 37,800 dalton의 band가 양적 증가를 보이는데 비해 membrane fraction 에서는 260,600 dalton의 band가 증가하였다. 이러한 결과로 성장 발달에 따라 고분자 량의 물질들이 이들 소뇌 부위에서 기여하였을 것으로 추정할 수 있었다. Immunocytochemistry에 의한 분석에서는 c-raf와 a-raf가 소뇌의 피질주위에서 조롱박 세포(Purkinje cell) 의 세포질 특히 핵 주변부위에서 강하게 검출되었으며 a-raf에 비해 c-raf가 더 강하게 나타났었다. 그리고 그 외에 Nucleus embolifornis의 큰 neuronal cell의 세포질 부위의 나타남을 볼 수 있었다. Immunoblot에 의한 분석에서는 crude와 cytosolic fraction에서 raf protein kinase의 존재를 확인할 수 있었으며, 이상의 결과들을 종합해 보았을 때 소뇌의 정상의 많은 신경세포(neuronal cell)에 raf protein kinase가 분포되어 있으며 이들이 정상의 cell에서 기능을 가질 것으로 추정된다.

a- and c-raf protein kinase in the brain of rat, the protein pattern of cerebellum during postnatal development of rat by polyacryamide gel electrophoresis, and the existence of c-raf protein kinase by using Western blotting method. The results were as follows: The cytoplasm of Purkinje cells was, in general, strongly labeled with the antibodies of a- and c-raf protein kinases in the cortex regions such as Pyramis cerebelli, Unula, Nodulus, Paraflocculus, and Flocculus. C-raf protein kinase appeared stronger immunoreactivity than a-raf protein kinase. In peripheral of cytoplasm of Nucleus emboliformis, A-raf Protein kinase was labeled markedly. During postnatal development, the protein of 38,000 dalton increased gradually in the cytosolic fraction of cerebellum, and the protein of 260,600 dalton appeared in the membrane fraction of cerebellum. By immunoblotting method, the protein band of 74,000 dalton was detected in crude and cytosolic fractions, but it was not exhibited in membrane fraction, In this fact, it was identified that a - and c-raf proteins were distributed throughout neuronal cells, especially in the Purkinje cells, in normal cerebellum cortex of rat. Also, this phenomenon was assumed that raf protein kinase in cytoplasm of neuronal cell had to do with a certain functional mechanism and signal transduction of neurotransmitter as Protein kinase C.

키워드

참고문헌

  1. Ann. Rev. Biochem. v.52 Cellular oncogenes and retrovirus Bishop, J.M.
  2. Nucl. Acids. Res. v.15 The complete coding sequence of the human a-raf-1 oncogne and transforming activity of a human a-raf carrying retrovirus Beck, T.W.;Huleihel, M.;Gunnell, M.;Bonner, T.I.;Rapp, U.R.
  3. Nucl. Acids Res. v.14 The complete coding seqeunce of he human raf oncogene and the correspondig strusture of c-raf-1 gene Bonner, T.I.;Oppermann, H.;Seeburg, P.;Kerby, S.B.;Gunnell, M.A.;Young, A.C.;Rapp, U.R.
  4. J. Virol. v.45 New mammalian transforming retrovirus: demonstration of a polyprotein gene product. Rapp, U.R.;Reynolds, F.H.;Stephenson, J.R.
  5. Mol. Cell. Biol. v.5 Structure and Biological activity of human honologs of the raf/mil oncogene Bonner, T.I.;Kerby, S.B.;Sutrave, P.;Gunnell, M.A.;Mark, C.;Rapp, U.R.
  6. Proc. natl. Acad. Sci. v.82 Detection of a raf-related and two other transforming DNA sequence in Human tumors maintained in nude mice Fukui, M.;Yamamoto, T.;Kawai, S.;Maruo, K.;Toyoshima, K.
  7. Proc. Natl. Acad. Aci. v.82 Molecular cloning of unactivated human oncogene, homologous to v-raf, from primary stomach cancer Shimuzu, K.;Nakatsu, Y.;Sekiguchi, M.;Hokamura, K.;Tanaka, M.;Sugimura, T.
  8. Proc. Natl. Acad. Aci. v.83 Actively transcribed genes in the raf oncegene group, located on the X chromosome in mouse and human Huebner, K.;ar-Rushdi, A.;Griffin, C.A.;Isobe, M.;Kozak, C.;Eemanuel, B.S.;Nafarajan, L.;Cleveland, J.L.;Bonner, T.I.;Goldsborough, M.D.;Croce, C.M.;Rapp, U.R.
  9. Mol. Cell. Biol. v.6 no.26 Characterization of murine a-raf, a new oncogene related ot the v-raf oncogene Huleihel, M.;Goldsborough, M.M.D.;Cleveland, J.L.;Gunnell, M.;Bonner, T.I.;Rapp, U.R.
  10. Proc. Natl Acad. Sci. v.83 pks, a raf related sequence in humans Mark, G.B.;Seeley, T.W.; Shows, T.B.;Mountx, J.D.
  11. Jpn. J. Cancer Res. v.77 Cystein-rich regions conserved in amino-terminal halves of raf gene famoly products and kinase C Ishikawa F.;Thakaku, F.;Nagao, M.;Sugimura, T.
  12. Science v.233 The complete Primary strcture of protein Kinase C-the major phorbol ester receptor Parker, P.J.;Stabel, S.;Waterfield, M.D.;Rhee, L.;Cousens, L.;Toffy, N.;Young, S.L;Chen, E.Ullrich, A.
  13. Science v.233 Multiple. distinct forms of bovine and human progein kinase C suggest diversity in cellular signaling pathways Coussens, I.;Parker, P.J.;Rhee, L.;Yang-Feng, T.L.;Chen E.;Waterfield, M.D.;Franke, U.;Ullrich, A.
  14. Celll v.46 Clonging and expression of multiple Protein Kinaes C cDNA Knoph, J.L.;Lee, M.H.;Sultzman, L.A.;Kirtz, R.W.;Loomis, C.R.lHewrck, R.M.;Bell, R.M.
  15. FEBS Lett. v.206 Teo types of compelementary DNAs of raf brain portein kinase C Ono, Y.;Kurokawa, T.;Fujii, T.;Kawahara, K.;Igarashi, K.;Kikkawa, U.;Rgita, K.;Nishizuka, Y.
  16. J. Biol. Chem. v.252 Studies on a cyclic nucleotide-independent protein kinase and its proenzyme in mammalian tissues Nishizuka, Y.
  17. J. Biol. Chem. v.252 Studies on a cyclic mucleotide-independent protein kinase and its proenzyme in mammalian tissues Takai, Y.;Kishimoto, A.;Inoue, M.;Nishizuka, Y.
  18. Mol. Biol. Biochem. Biophys. v.32 Three Multifunctional protein kinase in transmembrane control Nishizuka, Y.
  19. Mol. & Cell. Endorcrinol. v.35 Phospholipid-sensitive Ca++-dependent protein kinase : a major protein phosphorylation system Kuo, J.F.;Schatzman, R.C.;Turner, R.S.;Mazzei, G.J.
  20. Proc. Nail. Acad. Sci. v.82 Polyclonal antibodies to phospholipid/Ca++-dependent protein kinase and immunocytochemical localization of the enzyme in rat brain Girard, P.G.;Mazzei, G.J.;Wood, J.G.;Kuo, J.F.
  21. J. Biol. Chem. v.261 Immunological quantitation of phospholipid/Ca++-dependent protein knase and its fragment Girard, P.R.;Mazzei, G.J.;Kuo, J.F.
  22. J. Neurosci. v.6 Immuno-cytochemical localization of protein kinase C in identified meuronal compartments of rat brain Wood, J.G.;Girard, P.R.;Mazzei, G.J. Kuo, J.F.
  23. J. Biol. Chem. v.193 Protein measurement with the folin phenol reagent Lowry, O.H.;Rosebroyg, N.J.;Farr, A.L.;Rondoll, R.J.
  24. Nautre v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli, U.K
  25. Nature and Life v.17 Computer programs for estimation of molecular weight by electrophoretic method Choi, W.C.
  26. Proc. Natl. Acad. Sci. v.85 Phosphoipase Cc Ⅰand Ⅱ brain isozyme: immunohistochemical localization in neuronal system in rat brain Gerfen, C.R.;Choi, W.C.;Suh, P.B.;Rhee, S.G.
  27. J. Biol. Chem. v.84 immunocytochemical localization of calmodulin and a heat label calmodulin binding brain Wood, J.G.;Wallance, R.W.;Whitaker, J.N.;Cheung, W.Y.
  28. Proc. Natl. Acad. Sci. v.80 Structure and biological activity of v-raf, a unique oncogene transduced by retrovirus Rapp, U.R.;Goldsborough, M.D.;Mark, G.E.;Bonner, T.I.;Gorffen, J.; Reynolds, F.H.;Stephenson, J.
  29. J. Cell. Biochem. v.30 Interaction between raf nad myc oncogenes in transformation in vivo and in vitro Cleveleand, J.L.;Jansen, H.W.; Bister, K.;Fredrickson, T.N.;Morse Ⅲ, H.C.;Ihle, J.N.;Rapp, U.R.
  30. Ulrecht transformation by raf and myconcogenes Rapp, U.R.;Cleveland, J.L.;Storm, S.M.;Beck, T.W.;Heleichel, M.
  31. Role of raf and myc oncogenes in signal transdurction In genes and singal transcution in multistage,Crcinogenesis Heidecker, G.;Cleveland. J.L.:Beck, T.W.;Heleichel, M.;Kolch, W.;Storm, S.M.;Rapp, U.R.;N. Coburn(ed.)
  32. Cold Spting Harbpr Symposia Quentilative Biology v.Ⅲ raf family serine/threonine protein kinase in mitogen signal transduction Rapp, U.R.;Heidecker, G.;Huleihel, M.;Cleveland, J.L.;Choi, W.C.;Pawson, T.;Ihle, J.N.;Anderson, W.B.
  33. Adv. Cycl. Nucl. Prot. Phos. Res. v.19 Altered subcellelar distribution of progein kianse C ( aphorbol ester receptor) Anderson, W.B.;Estival, A.;Tapiovaara, H.;Gopalakristna, R.
  34. J. Biol. Chem. v.257 Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor promoting phorbol esters Castagna, M.;Takai, Y.;Kaibuchyi, K.;Kikkawa, V.;Nishizuka, Y.
  35. Science v.233 Strudies and perspectives of protein kinase c Nkshizuka, Y.
  36. Boichem v.25 Rapid microassay for protein kianse c translocation in swiss 3T3 cells Pelch, S.L.;Meier, K.E.;Krebs, E.G.
  37. Science v.225 turnover of inositol phospholipids and signal transduction Nishizuka, Y.
  38. Cell v.45 Protein kinase C activatied by diacylglycerol second messengers Bell, R.M.
  39. Cell v.33 Molecular cloning of gene sequences regulated by platelet derived growth factor Cochran, B.M.;Reffel, A.C.;Stiles, C.D.
  40. Craigiels neuroanatomy of the rat. Zemean, W.J.;Innes, R.M.
  41. Brain Res. v.90 Comparison fo postnatalCNS development between male and female rats Gregory, E.
  42. J. Comp. Neur. v.133 An electron microscopic study of the neuroglia during postnatal development of the rat cerebrum Caley, C.W.;Maxwell, D.S.
  43. J. Neurosci. v.6 PCCP-260 a Purkinje cell-specific cyclic AMP-regulated membrane phosphoprotein of molecular weight 260, 000 Walass, S.I.;Nairn, A.C.;Greengard, P.