메뉴 건너뛰기




Volumn 286, Issue 5444, 1999, Pages 1583-1587

Regulation of myosin phosphatase by a specific interaction with cGMP- dependent protein kinase Iα

Author keywords

[No Author keywords available]

Indexed keywords

CALCIUM; CYCLIC GMP; CYCLIC GMP DEPENDENT PROTEIN KINASE; LEUCINE; MYOSIN; MYOSIN LIGHT CHAIN KINASE; NITRIC OXIDE; PHOSPHATASE;

EID: 0033584786     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.286.5444.1583     Document Type: Article
Times cited : (458)

References (61)
  • 1
    • 0021894617 scopus 로고
    • K E. Kamm and J T. Stull, Annu Rev Pharmacol. Toxicol. 25, 593 (1985); D J Hartshorne, in Physiology of the Gastrointestinal Tract, D. R. Johnson, Ed. (Raven, New York, ed. 2, 1987), vol. 1, pp. 423-482; R. S. Filo, D. F. Bohr, J. C Ruegg, Science 147, 1581 (1965); A. P. Somlyo and B. Himpens, FASEB J. 3, 2266 (1989).
    • (1985) Annu Rev Pharmacol. Toxicol. , vol.25 , pp. 593
    • Kamm, K.E.1    Stull, J.T.2
  • 2
    • 0021894617 scopus 로고
    • D. R. Johnson, Ed. Raven, New York, ed. 2
    • K E. Kamm and J T. Stull, Annu Rev Pharmacol. Toxicol. 25, 593 (1985); D J Hartshorne, in Physiology of the Gastrointestinal Tract, D. R. Johnson, Ed. (Raven, New York, ed. 2, 1987), vol. 1, pp. 423-482; R. S. Filo, D. F. Bohr, J. C Ruegg, Science 147, 1581 (1965); A. P. Somlyo and B. Himpens, FASEB J. 3, 2266 (1989).
    • (1987) Physiology of the Gastrointestinal Tract , vol.1 , pp. 423-482
    • Hartshorne, D.J.1
  • 3
    • 0001658810 scopus 로고
    • K E. Kamm and J T. Stull, Annu Rev Pharmacol. Toxicol. 25, 593 (1985); D J Hartshorne, in Physiology of the Gastrointestinal Tract, D. R. Johnson, Ed. (Raven, New York, ed. 2, 1987), vol. 1, pp. 423-482; R. S. Filo, D. F. Bohr, J. C Ruegg, Science 147, 1581 (1965); A. P. Somlyo and B. Himpens, FASEB J. 3, 2266 (1989).
    • (1965) Science , vol.147 , pp. 1581
    • Filo, R.S.1    Bohr, D.F.2    Ruegg, J.C.3
  • 4
    • 0024459688 scopus 로고
    • K E. Kamm and J T. Stull, Annu Rev Pharmacol. Toxicol. 25, 593 (1985); D J Hartshorne, in Physiology of the Gastrointestinal Tract, D. R. Johnson, Ed. (Raven, New York, ed. 2, 1987), vol. 1, pp. 423-482; R. S. Filo, D. F. Bohr, J. C Ruegg, Science 147, 1581 (1965); A. P. Somlyo and B. Himpens, FASEB J. 3, 2266 (1989).
    • (1989) FASEB J. , vol.3 , pp. 2266
    • Somlyo, A.P.1    Himpens, B.2
  • 6
    • 0027049145 scopus 로고
    • D. Alessi, L. K. MacDougall, M. M. Sola, M. Ikebe, P. Cohen, Eur. J. Biochem. 210, 1023 (1992); K. Hirano, B. C. Phan, D. J. Hartshorne, J. Biol. Chem. 272, 3683 (1997); D. F. Johnson et al., Eur. J. Biochem. 239, 317 (1996); H. Shimizu et al., J. Biol. Chem 269, 30407 (1994).
    • (1992) Eur. J. Biochem. , vol.210 , pp. 1023
    • Alessi, D.1    MacDougall, L.K.2    Sola, M.M.3    Ikebe, M.4    Cohen, P.5
  • 7
    • 0031034680 scopus 로고    scopus 로고
    • D. Alessi, L. K. MacDougall, M. M. Sola, M. Ikebe, P. Cohen, Eur. J. Biochem. 210, 1023 (1992); K. Hirano, B. C. Phan, D. J. Hartshorne, J. Biol. Chem. 272, 3683 (1997); D. F. Johnson et al., Eur. J. Biochem. 239, 317 (1996); H. Shimizu et al., J. Biol. Chem 269, 30407 (1994).
    • (1997) J. Biol. Chem. , vol.272 , pp. 3683
    • Hirano, K.1    Phan, B.C.2    Hartshorne, D.J.3
  • 8
    • 0030036564 scopus 로고    scopus 로고
    • D. Alessi, L. K. MacDougall, M. M. Sola, M. Ikebe, P. Cohen, Eur. J. Biochem. 210, 1023 (1992); K. Hirano, B. C. Phan, D. J. Hartshorne, J. Biol. Chem. 272, 3683 (1997); D. F. Johnson et al., Eur. J. Biochem. 239, 317 (1996); H. Shimizu et al., J. Biol. Chem 269, 30407 (1994).
    • (1996) Eur. J. Biochem. , vol.239 , pp. 317
    • Johnson, D.F.1
  • 9
    • 0027972853 scopus 로고
    • D. Alessi, L. K. MacDougall, M. M. Sola, M. Ikebe, P. Cohen, Eur. J. Biochem. 210, 1023 (1992); K. Hirano, B. C. Phan, D. J. Hartshorne, J. Biol. Chem. 272, 3683 (1997); D. F. Johnson et al., Eur. J. Biochem. 239, 317 (1996); H. Shimizu et al., J. Biol. Chem 269, 30407 (1994).
    • (1994) J. Biol. Chem , vol.269 , pp. 30407
    • Shimizu, H.1
  • 10
    • 0023162273 scopus 로고
    • B. Bradley and K. G. Morgan, J. Physiol (London) 385, 437 (1987); T. Kitazawa, B. D. Gaylinn, G. H. Denney, A P. Somlyo, J. Biol. Chem. 266, 1708 (1991); M. Masuo, S. Reardon, M Ikebe, T. Kitazawa, J. Gen. Physiol. 104, 265 (1994), M. Cui Gong et al., J. Biol. Chem. 267, 21492 (1992), K. Kimura et al , Science 273, 245 (1996).
    • (1987) J. Physiol (London) , vol.385 , pp. 437
    • Bradley, B.1    Morgan, K.G.2
  • 11
    • 0025978177 scopus 로고
    • B. Bradley and K. G. Morgan, J. Physiol (London) 385, 437 (1987); T. Kitazawa, B. D. Gaylinn, G. H. Denney, A P. Somlyo, J. Biol. Chem. 266, 1708 (1991); M. Masuo, S. Reardon, M Ikebe, T. Kitazawa, J. Gen. Physiol. 104, 265 (1994), M. Cui Gong et al., J. Biol. Chem. 267, 21492 (1992), K. Kimura et al , Science 273, 245 (1996).
    • (1991) J. Biol. Chem. , vol.266 , pp. 1708
    • Kitazawa, T.1    Gaylinn, B.D.2    Denney, G.H.3    Somlyo, A.P.4
  • 12
    • 0028168041 scopus 로고
    • B. Bradley and K. G. Morgan, J. Physiol (London) 385, 437 (1987); T. Kitazawa, B. D. Gaylinn, G. H. Denney, A P. Somlyo, J. Biol. Chem. 266, 1708 (1991); M. Masuo, S. Reardon, M Ikebe, T. Kitazawa, J. Gen. Physiol. 104, 265 (1994), M. Cui Gong et al., J. Biol. Chem. 267, 21492 (1992), K. Kimura et al , Science 273, 245 (1996).
    • (1994) J. Gen. Physiol. , vol.104 , pp. 265
    • Masuo, M.1    Reardon, S.2    Ikebe, M.3    Kitazawa, T.4
  • 13
    • 0026806485 scopus 로고
    • B. Bradley and K. G. Morgan, J. Physiol (London) 385, 437 (1987); T. Kitazawa, B. D. Gaylinn, G. H. Denney, A P. Somlyo, J. Biol. Chem. 266, 1708 (1991); M. Masuo, S. Reardon, M Ikebe, T. Kitazawa, J. Gen. Physiol. 104, 265 (1994), M. Cui Gong et al., J. Biol. Chem. 267, 21492 (1992), K. Kimura et al , Science 273, 245 (1996).
    • (1992) J. Biol. Chem. , vol.267 , pp. 21492
    • Cui Gong, M.1
  • 14
    • 9444242736 scopus 로고    scopus 로고
    • B. Bradley and K. G. Morgan, J. Physiol (London) 385, 437 (1987); T. Kitazawa, B. D. Gaylinn, G. H. Denney, A P. Somlyo, J. Biol. Chem. 266, 1708 (1991); M. Masuo, S. Reardon, M Ikebe, T. Kitazawa, J. Gen. Physiol. 104, 265 (1994), M. Cui Gong et al., J. Biol. Chem. 267, 21492 (1992), K. Kimura et al , Science 273, 245 (1996).
    • (1996) Science , vol.273 , pp. 245
    • Kimura, K.1
  • 15
    • 0032101741 scopus 로고    scopus 로고
    • A. Pfeifer et al , EMBO J. 17, 3045 (1998); T. M. Lincoln, in Cyclic GMP: Biochemistry, Physiology and Pathophysiology (R. G. Landes Company, Austin, TX, 1994), pp. 97-115; S. M Lohmann, A. B. Vaandrager, A. Smolenski, U Walter, H. R. DeJonge, Trends Biochem. Sci. 22, 307 (1997).
    • (1998) EMBO J. , vol.17 , pp. 3045
    • Pfeifer, A.1
  • 16
    • 0032101741 scopus 로고    scopus 로고
    • R. G. Landes Company, Austin, TX
    • A. Pfeifer et al , EMBO J. 17, 3045 (1998); T. M. Lincoln, in Cyclic GMP: Biochemistry, Physiology and Pathophysiology (R. G. Landes Company, Austin, TX, 1994), pp. 97-115; S. M Lohmann, A. B. Vaandrager, A. Smolenski, U Walter, H. R. DeJonge, Trends Biochem. Sci. 22, 307 (1997).
    • (1994) Cyclic GMP: Biochemistry, Physiology and Pathophysiology , pp. 97-115
    • Lincoln, T.M.1
  • 18
    • 0021646397 scopus 로고
    • J. P. Morgan and K. G. Morgan, J. Physiol. (London) 357, 539 (1984); T. M. Lincoln and T. L. Cornwell, FASEB J. 7, 328 (1993); T. L. Cornwell and T. M. Lincoln, J. Biol. Chem. 264, 1146 (1989); X. Wu, A. V. Somlyo, A. P. Somlyo, Biochem. Biophys. Res Commun. 220, 658 (1996); M. R. Lee, L. Li, T. Kitazawa, J. Biol. Chem. 272, 5063 (1997).
    • (1984) J. Physiol. (London) , vol.357 , pp. 539
    • Morgan, J.P.1    Morgan, K.G.2
  • 19
    • 0027404913 scopus 로고
    • J. P. Morgan and K. G. Morgan, J. Physiol. (London) 357, 539 (1984); T. M. Lincoln and T. L. Cornwell, FASEB J. 7, 328 (1993); T. L. Cornwell and T. M. Lincoln, J. Biol. Chem. 264, 1146 (1989); X. Wu, A. V. Somlyo, A. P. Somlyo, Biochem. Biophys. Res Commun. 220, 658 (1996); M. R. Lee, L. Li, T. Kitazawa, J. Biol. Chem. 272, 5063 (1997).
    • (1993) FASEB J. , vol.7 , pp. 328
    • Lincoln, T.M.1    Cornwell, T.L.2
  • 20
    • 0024476861 scopus 로고
    • J. P. Morgan and K. G. Morgan, J. Physiol. (London) 357, 539 (1984); T. M. Lincoln and T. L. Cornwell, FASEB J. 7, 328 (1993); T. L. Cornwell and T. M. Lincoln, J. Biol. Chem. 264, 1146 (1989); X. Wu, A. V. Somlyo, A. P. Somlyo, Biochem. Biophys. Res Commun. 220, 658 (1996); M. R. Lee, L. Li, T. Kitazawa, J. Biol. Chem. 272, 5063 (1997).
    • (1989) J. Biol. Chem. , vol.264 , pp. 1146
    • Cornwell, T.L.1    Lincoln, T.M.2
  • 21
    • 0029892621 scopus 로고    scopus 로고
    • J. P. Morgan and K. G. Morgan, J. Physiol. (London) 357, 539 (1984); T. M. Lincoln and T. L. Cornwell, FASEB J. 7, 328 (1993); T. L. Cornwell and T. M. Lincoln, J. Biol. Chem. 264, 1146 (1989); X. Wu, A. V. Somlyo, A. P. Somlyo, Biochem. Biophys. Res Commun. 220, 658 (1996); M. R. Lee, L. Li, T. Kitazawa, J. Biol. Chem. 272, 5063 (1997).
    • (1996) Biochem. Biophys. Res Commun. , vol.220 , pp. 658
    • Wu, X.1    Somlyo, A.V.2    Somlyo, A.P.3
  • 22
    • 0031057516 scopus 로고    scopus 로고
    • J. P. Morgan and K. G. Morgan, J. Physiol. (London) 357, 539 (1984); T. M. Lincoln and T. L. Cornwell, FASEB J. 7, 328 (1993); T. L. Cornwell and T. M. Lincoln, J. Biol. Chem. 264, 1146 (1989); X. Wu, A. V. Somlyo, A. P. Somlyo, Biochem. Biophys. Res Commun. 220, 658 (1996); M. R. Lee, L. Li, T. Kitazawa, J. Biol. Chem. 272, 5063 (1997).
    • (1997) J. Biol. Chem. , vol.272 , pp. 5063
    • Lee, M.R.1    Li, L.2    Kitazawa, T.3
  • 23
    • 0031457622 scopus 로고    scopus 로고
    • T. Pawson and J. D. Scott, Science 278, 2075 (1997); J. D. Scott and M L. DellAcqua, J. Biol Chem 272, 12881 (1997); J. D. Scott and M. C. Faux, Trends Biochem. Sci 21, 312 (1996); D. Mochly-Rosen, Science 268, 247 (1995); C. S. Rubin, Biochim. Biophys Acta 1224, 467 (1994); P. Cohen and M J. Hubbard, Trends Biochem. Sci. 18, 172 (1993).
    • (1997) Science , vol.278 , pp. 2075
    • Pawson, T.1    Scott, J.D.2
  • 24
    • 0030914913 scopus 로고    scopus 로고
    • T. Pawson and J. D. Scott, Science 278, 2075 (1997); J. D. Scott and M L. DellAcqua, J. Biol Chem 272, 12881 (1997); J. D. Scott and M. C. Faux, Trends Biochem. Sci 21, 312 (1996); D. Mochly-Rosen, Science 268, 247 (1995); C. S. Rubin, Biochim. Biophys Acta 1224, 467 (1994); P. Cohen and M J. Hubbard, Trends Biochem. Sci. 18, 172 (1993).
    • (1997) J. Biol Chem , vol.272 , pp. 12881
    • Scott, J.D.1    DellAcqua, M.L.2
  • 25
    • 0030219389 scopus 로고    scopus 로고
    • T. Pawson and J. D. Scott, Science 278, 2075 (1997); J. D. Scott and M L. DellAcqua, J. Biol Chem 272, 12881 (1997); J. D. Scott and M. C. Faux, Trends Biochem. Sci 21, 312 (1996); D. Mochly-Rosen, Science 268, 247 (1995); C. S. Rubin, Biochim. Biophys Acta 1224, 467 (1994); P. Cohen and M J. Hubbard, Trends Biochem. Sci. 18, 172 (1993).
    • (1996) Trends Biochem. Sci , vol.21 , pp. 312
    • Scott, J.D.1    Faux, M.C.2
  • 26
    • 0028938152 scopus 로고
    • T. Pawson and J. D. Scott, Science 278, 2075 (1997); J. D. Scott and M L. DellAcqua, J. Biol Chem 272, 12881 (1997); J. D. Scott and M. C. Faux, Trends Biochem. Sci 21, 312 (1996); D. Mochly-Rosen, Science 268, 247 (1995); C. S. Rubin, Biochim. Biophys Acta 1224, 467 (1994); P. Cohen and M J. Hubbard, Trends Biochem. Sci. 18, 172 (1993).
    • (1995) Science , vol.268 , pp. 247
    • Mochly-Rosen, D.1
  • 27
    • 0028588997 scopus 로고
    • T. Pawson and J. D. Scott, Science 278, 2075 (1997); J. D. Scott and M L. DellAcqua, J. Biol Chem 272, 12881 (1997); J. D. Scott and M. C. Faux, Trends Biochem. Sci 21, 312 (1996); D. Mochly-Rosen, Science 268, 247 (1995); C. S. Rubin, Biochim. Biophys Acta 1224, 467 (1994); P. Cohen and M J. Hubbard, Trends Biochem. Sci. 18, 172 (1993).
    • (1994) Biochim. Biophys Acta , vol.1224 , pp. 467
    • Rubin, C.S.1
  • 28
    • 0027277098 scopus 로고
    • T. Pawson and J. D. Scott, Science 278, 2075 (1997); J. D. Scott and M L. DellAcqua, J. Biol Chem 272, 12881 (1997); J. D. Scott and M. C. Faux, Trends Biochem. Sci 21, 312 (1996); D. Mochly-Rosen, Science 268, 247 (1995); C. S. Rubin, Biochim. Biophys Acta 1224, 467 (1994); P. Cohen and M J. Hubbard, Trends Biochem. Sci. 18, 172 (1993).
    • (1993) Trends Biochem. Sci. , vol.18 , pp. 172
    • Cohen, P.1    Hubbard, M.J.2
  • 29
    • 0029887701 scopus 로고    scopus 로고
    • T. M. Klauck et al., Science 271, 1589 (1996); K. Choi, B. Satterberg, D. M Lyons, E. A. Elion, Cell 78, 499 (1994).
    • (1996) Science , vol.271 , pp. 1589
    • Klauck, T.M.1
  • 31
    • 13044305938 scopus 로고    scopus 로고
    • note
    • The full-length coding region of bovine cGKIα (10) was cloned into the Gal4 DNA binding domain vector PC97 (11). cGKPC97 and a human activated T cell library (cloned into the Gal4 DNA activating domain vector PC86, gift of M. Vidal) were transformed into S. cerevisiae strain MaV103 (MATa, leu2-3, 112 trp1-901 his3△200 ade2-101 gal4△ gal80△ SPAL10-:URA3 GAL1::LacZ GAL1::HIS3@Lys2 Can1R cyh2R) using the lithium acetate method (12). Polypeptides interacting with CGKPC97 were detected by their ability to activate transcription of HIS3 and LacZ reporter genes PC86 plasmids from HIS+, LacZ+ colonies were isolated, and the library cDNA insert was sequenced. To confirm the interaction of AL9 with cGKIα, individual plasmids were reintroduced into 5. cerevisiae strain Y190 (MAT a, ura3-52, his3-200, Lys2-801, ade2-101, trp1-901, leu2-3, 112, gal4△, gal80△, cyhr2, LYS2::GAl.1UAS-HIS3TATA-HIS3, URA3::GAL1UAS-GAL1TATA-lacZ) (Clontech) by the lithium acetate method, and reporter activation was assayed as above.
  • 33
    • 0026639625 scopus 로고
    • P Chevray and D Nathans, Proc. Natl. Acad. Sci. U.S.A. 89, 5789 (1992); M. Vidal, R. K Brachmann, A. Fattaey, E. Harlow, J. D. Boeke, Proc. Natl. Acad. Sci U.S.A. 93, 10315 (1996).
    • (1992) Proc. Natl. Acad. Sci. U.S.A. , vol.89 , pp. 5789
    • Chevray, P.1    Nathans, D.2
  • 37
    • 13044260289 scopus 로고    scopus 로고
    • note
    • 68-667 mutants. Pst I sites were engineered at amino acids 4 and 68 with degenerate polymerase chain reaction primers. The Pst I fragment was excised, and the COOH-terminal cGKIa was religated and transformed into yeast strain Y190. The proper reading frame was verified by DNA sequencing. Expression of the COOH-terminal construct in PC97 was confirmed by a protein immunoblot of the Y190 lysate, using the rabbit polyclonal Gal4 DNA-binding domain antibody (Upstate Biotechnology, Lake Placid, NY).
  • 38
    • 0025925410 scopus 로고
    • R. A. Atkinson, V. Saudek, J. P. Huggins, J. T. Pelton, Biochemistry 30, 9387 (1991); W. H. Landschulz, P. F. Johnson, S. L. McKnight, Science 240, 1759 (1988), W M Landschulz, P. F. Johnson, S. L. McKnight, Science 243, 1681 (1989); R Turner and R. Tjian, Science 243, 1689 (1989); P. B. Harbury, T. Zhang, P. S. Kim, T. Alber, Science 262, 1401 (1993)
    • (1991) Biochemistry , vol.30 , pp. 9387
    • Atkinson, R.A.1    Saudek, V.2    Huggins, J.P.3    Pelton, J.T.4
  • 39
    • 0024295767 scopus 로고
    • R. A. Atkinson, V. Saudek, J. P. Huggins, J. T. Pelton, Biochemistry 30, 9387 (1991); W. H. Landschulz, P. F. Johnson, S. L. McKnight, Science 240, 1759 (1988), W M Landschulz, P. F. Johnson, S. L. McKnight, Science 243, 1681 (1989); R Turner and R. Tjian, Science 243, 1689 (1989); P. B. Harbury, T. Zhang, P. S. Kim, T. Alber, Science 262, 1401 (1993)
    • (1988) Science , vol.240 , pp. 1759
    • Landschulz, W.H.1    Johnson, P.F.2    McKnight, S.L.3
  • 40
    • 0024518919 scopus 로고
    • R. A. Atkinson, V. Saudek, J. P. Huggins, J. T. Pelton, Biochemistry 30, 9387 (1991); W. H. Landschulz, P. F. Johnson, S. L. McKnight, Science 240, 1759 (1988), W M Landschulz, P. F. Johnson, S. L. McKnight, Science 243, 1681 (1989); R Turner and R. Tjian, Science 243, 1689 (1989); P. B. Harbury, T. Zhang, P. S. Kim, T. Alber, Science 262, 1401 (1993)
    • (1989) Science , vol.243 , pp. 1681
    • Landschulz, W.M.1    Johnson, P.F.2    McKnight, S.L.3
  • 41
    • 0024535960 scopus 로고
    • R. A. Atkinson, V. Saudek, J. P. Huggins, J. T. Pelton, Biochemistry 30, 9387 (1991); W. H. Landschulz, P. F. Johnson, S. L. McKnight, Science 240, 1759 (1988), W M Landschulz, P. F. Johnson, S. L. McKnight, Science 243, 1681 (1989); R Turner and R. Tjian, Science 243, 1689 (1989); P. B. Harbury, T. Zhang, P. S. Kim, T. Alber, Science 262, 1401 (1993)
    • (1989) Science , vol.243 , pp. 1689
    • Turner, R.1    Tjian, R.2
  • 42
    • 0027756896 scopus 로고
    • R. A. Atkinson, V. Saudek, J. P. Huggins, J. T. Pelton, Biochemistry 30, 9387 (1991); W. H. Landschulz, P. F. Johnson, S. L. McKnight, Science 240, 1759 (1988), W M Landschulz, P. F. Johnson, S. L. McKnight, Science 243, 1681 (1989); R Turner and R. Tjian, Science 243, 1689 (1989); P. B. Harbury, T. Zhang, P. S. Kim, T. Alber, Science 262, 1401 (1993)
    • (1993) Science , vol.262 , pp. 1401
    • Harbury, P.B.1    Zhang, T.2    Kim, P.S.3    Alber, T.4
  • 43
    • 13044279595 scopus 로고    scopus 로고
    • note
    • 2, 2 mM EDTA, 2 mg/ml N-dodecyl-B-maltoside, 0.4 mg/ml cholesteryl hemisuccinate, 0.6 M NaCl, 10 mM Na Molybdate, 1 mM phenylmethylsulfonyl fluoride (PMSF), 10 μg/ml chymostatin, 200 μg/ml aprotinin, 50 μg/ml leupeptin] and incubated for 1 hour at room temperature. Lysates were microfuged for 15 mm at 4°C, and the supernatant was incubated with 100 μI of GST-fusion protein beads overnight, followed by washing in RIPA buffer containing 1% NP40, and boiling for 5 min in SDS sample buffer. Associated proteins were resolved by SDS-PAGE, transferred to nitrocellulose, and immunoblotted with either rabbit anti-human cGPK-CT (Upstate Biotechnology, Lake Placid, NY) or rabbit anti-MBS antibody (Berkelely Antibody Company). The membranes were developed with ECL (Amersham Life Science).
  • 45
    • 0028358761 scopus 로고
    • Fluorescence spectroscopy was performed as previously described for the binding of cGK and vimentin (L. A. MacMillan-Crow and T. M. Lincoln, Biochemistry 33, 8035 (1994)].
    • (1994) Biochemistry , vol.33 , pp. 8035
    • MacMillan-Crow, L.A.1    Lincoln, T.M.2
  • 47
    • 13044280878 scopus 로고    scopus 로고
    • note
    • 4,5A were each cotransformed with AL9 in yeast strain Y190, and reporter gene activation assayed as in (9).
  • 48
    • 13044304651 scopus 로고    scopus 로고
    • note
    • 2. 2.5 mM EDTA, 1% Triton X and protease inhibitors as in buffer A). The lysate was incubated for 1 hour at room temperature, microcentrifuged 5 s, and the supernatant precleared with 12-5 μg rabbit IgG followed by protein A beads The precleared supernatant was incubated with either rabbit nonimmune IgG, or rabbit polyclonal anti-MBS (Berkeley Antibody Company) overnight, followed by harvest with protein A beads. Equal amounts of rabbit nonimmune and anti-MBS antibodies were added, and verified by Ponceau staining. SDS-PAGE and cGK immunoblots were performed as above.
  • 49
    • 13044274265 scopus 로고    scopus 로고
    • note
    • 32P-myosin light chains were used as substrate in place of phosphorylase a, as previously described (4), and similar results were obtained Data are presented as mean ± standard error.
  • 50
    • 0024517280 scopus 로고
    • H. Ishihara et al , Biochem. Biophys. Res. Commun. 159, 871 (1989); D. L Brautigan and C. L Shriner, Methods Enzymol. 159, 339 (1988); P. Cohen, Methods Enzymol 201, 389 (1991).
    • (1989) Biochem. Biophys. Res. Commun. , vol.159 , pp. 871
    • Ishihara, H.1
  • 51
    • 0023761334 scopus 로고
    • H. Ishihara et al , Biochem. Biophys. Res. Commun. 159, 871 (1989); D. L Brautigan and C. L Shriner, Methods Enzymol. 159, 339 (1988); P. Cohen, Methods Enzymol 201, 389 (1991).
    • (1988) Methods Enzymol. , vol.159 , pp. 339
    • Brautigan, D.L.1    Shriner, C.L.2
  • 52
    • 0025998844 scopus 로고
    • H. Ishihara et al , Biochem. Biophys. Res. Commun. 159, 871 (1989); D. L Brautigan and C. L Shriner, Methods Enzymol. 159, 339 (1988); P. Cohen, Methods Enzymol 201, 389 (1991).
    • (1991) Methods Enzymol , vol.201 , pp. 389
    • Cohen, P.1
  • 53
    • 0017387814 scopus 로고
    • 32P]ATP, for 15 min at 30°C SDS-PAGE was performed as above cGK-CA was prepared by incubating 50 μg of bovine cGKIα [purified as in T. M. Lincoln, W. L Dills, J. D. Corbin, J. Biol. Chem. 252, 4269 (1977)] with 1 μg of trypsin for 3 min at 30°C The reaction was terminated by the addition of 5 μg of soybean trypsin inhibitor. Image analysis of gel bands was performed using Scion Image software, and data are presented as mean ± SE
    • (1977) J. Biol. Chem. , vol.252 , pp. 4269
    • Lincoln, T.M.1    Dills, W.L.2    Corbin, J.D.3
  • 55
    • 0019321405 scopus 로고
    • C. E. Monken and G. N.Gill, J. Biol. Chem. 255, 7067 (1980); W. G. Heil, W Landgraf, F. Hofmann, Eur. J Biochem 168, 117 (1987).
    • (1980) J. Biol. Chem. , vol.255 , pp. 7067
    • Monken, C.E.1    Gill, G.N.2
  • 57
    • 13044264559 scopus 로고    scopus 로고
    • note
    • Human saphenous vein smooth muscle cells of passage 2-4 were grown on coverslips, fixed with 3% paraformaldehyde then permeabilized with 0.3% Triton X 100. For preservation of stress fiber architecture prior to fixation, cells were washed on ice with PBS for 1 min and permeabilized with 0.3% Triton X in 50 mM tris (pH 7.4) with 0.5 mM PMSF, 1 μg/ml leupeptin, 1 μg/ml aprotinin, and 1 μg/ml pepstatin on ice for 1 min The cells were then washed with PBS and fixed as above. For both protocols, cells were blocked with 10% donkey serum in PBS for 1 hour at 37°C, and washed with PBS. Primary antibody mixtures were rabbit polyclonal anti-MBS (1/125), or goat polyclonal anti-cGK (1/250). Secondary antibodies were donkey anti-rabbit IgG-conjugated Cy3 (Amersham Life Science) (1/800) and donkey anti-goat IgG-conjugated fluorescein isothiocyanate (Chemicon International Inc) (1/100). Following incubation with secondary antibody, the coverslips were washed with PBS and mounted in Slow Fade (Molecular Probes).
  • 59
    • 0023746769 scopus 로고
    • 1-59PCI by electroporation. Cells were arrested in DMEM supplemented with insulin, ascorbic acid and transferrin, then stimulated with 2 μM U46619 for 1 minute with or without 20-min pretreatment with 1 mM 8-Br-cGMP Following treatment, cells were precipitated with TCA, washed with acetone, and protein was subjected to glycerolurea electrophoresis [D. A. Taylor and J. T. Stull. J. Biol. Chem. 263, 14456 (1988)]. The nonphosphorylated and phosphorylated forms of myosin light chain were detected by immunoblotting with monoclonal anti-myosin light chain antibody (Sigma, clone MY-21). Protein bands were analyzed by densitometry Statistical analysis was performed using Student-Newman-Keuls method.
    • (1988) J. Biol. Chem. , vol.263 , pp. 14456
    • Taylor, D.A.1    Stull, J.T.2
  • 61
    • 13044256493 scopus 로고    scopus 로고
    • note
    • We thank D. L. Brautigan for generosity with reagents and for several helpful discussions, M. Vidal for reagents and advice regarding the two hybrid screens, L. J Moss for help with confocal microscopy, and L. A. MacMillan-Crow for assistance in performing the fluorescence spectroscopy. Supported in part by NIH grants HL09330 (H.K.S.) and HL5S309 (M.E.M.). M.E.M is an Established Investigator of the American Heart Association.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.