Targeting the receptor-G(q) interface to inhibit in vivo pressure overload myocardial hypertrophy

Science

Volumn 280, Issue 5363, 1998, Pages 574-577

  Akhter, Shahab A ^a   Luttrell, Louis M ^a   Rockman, Howard A ^a   Iaccarino, Guido ^a   Lefkowitz, Robert J ^a   Koch, Walter J ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

GUANINE NUCLEOTIDE BINDING PROTEIN; PROTEIN SUBUNIT; ALPHA ADRENERGIC RECEPTOR; ANGIOTENSIN II; ATRIAL NATRIURETIC FACTOR; DIACYLGLYCEROL; INOSITOL PHOSPHATE; MITOGEN ACTIVATED PROTEIN KINASE 1; PEPTIDE FRAGMENT; PHENYLEPHRINE;

ARTICLE; CONTROLLED STUDY; HEART VENTRICLE HYPERTROPHY; HUMAN; HUMAN CELL; HUMAN TISSUE; INHIBITION KINETICS; PATHOPHYSIOLOGY; PRESSURE GRADIENT; PRIORITY JOURNAL; PROTEIN BINDING; RECEPTOR BINDING; SARCOLEMMA; SIGNAL TRANSDUCTION; ANIMAL; CELL STRAIN COS1; DRUG ANTAGONISM; ENZYME ACTIVATION; GENE EXPRESSION REGULATION; GENE TARGETING; GENETIC TRANSFECTION; GENETICS; HEART LEFT VENTRICLE HYPERTROPHY; HEART MUSCLE; HEART VENTRICLE PRESSURE; METABOLISM; MOUSE; TRANSGENE; TRANSGENIC MOUSE;

ANGIOTENSIN II; ANIMALS; ATRIAL NATRIURETIC FACTOR; COS CELLS; DIGLYCERIDES; ENZYME ACTIVATION; GENE EXPRESSION REGULATION; GENE TARGETING; GTP-BINDING PROTEINS; HYPERTROPHY, LEFT VENTRICULAR; INOSITOL PHOSPHATES; MICE; MICE, TRANSGENIC; MITOGEN-ACTIVATED PROTEIN KINASE 1; MYOCARDIUM; PEPTIDE FRAGMENTS; PHENYLEPHRINE; RECEPTORS, ADRENERGIC, ALPHA; SIGNAL TRANSDUCTION; TRANSFECTION; TRANSGENES; VENTRICULAR PRESSURE;

EID: 0032562289     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.280.5363.574     Document Type: Article

References (32)

1. K. R. Chien, K. U. Knowlton, H. Zhu, S. Chien, FASEB J. 5, 3037 (1991).
2. P. Simpson, J. Clin. Invest. 72, 732 (1983); H. E. Shubeita et al., J. Biol. Chem. 265, 20555 (1990); H. Ito et al., J. Clin. Invest. 92, 398 (1993); J. Sadoshima, Y. Xu, H. S. Slayter, S. Izumo, Cell 75, 977 (1993).
3. P. Simpson, J. Clin. Invest. 72, 732 (1983); H. E. Shubeita et al., J. Biol. Chem. 265, 20555 (1990); H. Ito et al., J. Clin. Invest. 92, 398 (1993); J. Sadoshima, Y. Xu, H. S. Slayter, S. Izumo, Cell 75, 977 (1993).
4. P. Simpson, J. Clin. Invest. 72, 732 (1983); H. E. Shubeita et al., J. Biol. Chem. 265, 20555 (1990); H. Ito et al., J. Clin. Invest. 92, 398 (1993); J. Sadoshima, Y. Xu, H. S. Slayter, S. Izumo, Cell 75, 977 (1993).
5. P. Simpson, J. Clin. Invest. 72, 732 (1983); H. E. Shubeita et al., J. Biol. Chem. 265, 20555 (1990); H. Ito et al., J. Clin. Invest. 92, 398 (1993); J. Sadoshima, Y. Xu, H. S. Slayter, S. Izumo, Cell 75, 977 (1993).
6. H. A. Rockman, S. P. Wachhorst, L. Mao, J. Ross Jr., Am. J. Physiol. 266, H2468 (1994); H. Ito et al., Circulation 89, 2198 (1994).
7. H. A. Rockman, S. P. Wachhorst, L. Mao, J. Ross Jr., Am. J. Physiol. 266, H2468 (1994); H. Ito et al., Circulation 89, 2198 (1994).
8. C. A. Milano et al., Proc. Natl. Acad. Sci. U.S.A. 91, 10109 (1994).
9. L. Hein et al., ibid. 94, 6391 (1997).
10. D. D. D'Angelo et al., ibid., p. 8121.
11. E. J. Neer, Cell 80, 249 (1995).
12. L. M. Luttrell, J. Ostrowski, S. Cotecchia, H. Kendall, R. J. Lefkowitz, Science 259, 1453 (1993).
13. qI-SV-40. The transgene was then linearized and purified before pronuclear injections done by the Duke Comprehensive Cancer Center Transgenic Facility. Offspring were screened by Southern (DNA) blot analysis with a probe to the SV-40 sequences (4, 10, 20, 21). Institutional Review Board approval for all mouse experiments was obtained from each institution involved.
14. W. J. Koch, C. A. Milano, R. J. Lefkowitz, Circ. Res. 78, 511 (1996).
15. S. A. Akhter and W. J. Koch, unpublished data.
16. 32P-labeled diacylglycerol was isolated by silica gel thin-layer chromatography as described (4) and quantified with a Phosphorlmager (Molecular Dynamics). Diacylglycerol content is expressed as picomoles of diacylglycerol per nanomole of lipid phosphate (4).
17. 32P]ATP (20 μCi/ ml) and incubated at room temperature for 30 min. The reactions were quenched with 40 μl of 2× Laemmli buffer, and 30 μl of each reaction was electrophoresed through a 4 to 20% polyacrylamide:tris-glycine gel. Phosphorylated MBP on dried gels was quantified with a Phosphorlmager (Molecular Dynamics).
18. M. A. Bogoyevitch, A. Clerk, P. H. Sugden, Biochem. J. 309, 437 (1995).
19. Crude myocardial membranes were prepared as described (20, 21). Membrane proteins (20 to 30 μg) were incubated for 15 min at 37°C with [α-32P]ATP under basal conditions or with progressive doses of isoproterenol or 10 mM NaF, and cAMP was quantitated (20, 21).
20. 2.
21. H. A. Rockman et al., Proc. Natl. Acad. Sci. U.S.A. 88, 8277 (1991); K. U. Knowlton et al., J. Clin. Invest. 96, 1311 (1995); D.-J. Choi, W. J. Koch, J. J. Hunter, H. A. Rockman, J. Biol. Chem. 272, 17223 (1997).
22. H. A. Rockman et al., Proc. Natl. Acad. Sci. U.S.A. 88, 8277 (1991); K. U. Knowlton et al., J. Clin. Invest. 96, 1311 (1995); D.-J. Choi, W. J. Koch, J. J. Hunter, H. A. Rockman, J. Biol. Chem. 272, 17223 (1997).
23. H. A. Rockman et al., Proc. Natl. Acad. Sci. U.S.A. 88, 8277 (1991); K. U. Knowlton et al., J. Clin. Invest. 96, 1311 (1995); D.-J. Choi, W. J. Koch, J. J. Hunter, H. A. Rockman, J. Biol. Chem. 272, 17223 (1997).
24. V. J. LaMorte et al., J. Biol. Chem. 269, 13490 (1994).
25. S. Sakai et al., Nature 384, 353 (1996).
26. C. A. Milano et al., Science 264, 582 (1994).
27. W. J. Koch et al., ibid. 268, 1350 (1995).
28. All cDNAs were subcloned by standard methods into the eukaryotic expression vector pRK5 (8). The cDNA fragments were obtained by using standard polymerase chain reaction conditions, and sequences were confirmed by dideoxynucleotide sequencing (8).
29. 5 cells per well) the day before transfection and then transfected by using DEAE-dextran (8) and a total of 1 to 2 μg of DNA per well. Assays were performed 48 hours later.
30. 125I-labeled protein A (Amersham) or horseradish peroxidase-conjugated donkey antibody to rabbit immunoglobulin G (Jackson Laboratories) was used for detection.
31. Ventricular tissue was separated from the atria under a dissecting microscope. Total ventricular RNA was extracted with RNAzol (Biotecx) as described (20, 21).
32. We thank C. Bock at the Duke University Transgenic Mouse Facility for doing microinjections, J. Crosby and S. Duncan for technical assistance, and A. Eckhart for helpful discussions. Supported in part by NIH grants HL-16037 (R.J.L) and HL-03041 (H.A.R.), National Research Service Award HL-09436 (S.A.A), and a Grant-in-Aid from the North Carolina affiliate of the American Heart Association (W.J.K.).