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- Leroy, Marie-Josephe, et al.
(författare)
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Characterization of two recombinant PDE3 (cGMP-inhibited cyclic nucleotide phosphodiesterase) isoforms, RcGIP1 and HcGIP2, expressed in NIH 3006 murine fibroblasts and Sf9 insect cells
- 1996
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Ingår i: Biochemistry. - : American Chemical Society (ACS). - 0006-2960 .- 1520-4995. ; 35:31, s. 10194-10202
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Tidskriftsartikel (refereegranskat)abstract
- cDNAs encoding PDE3 [cGMP-inhibited cyclic nucleotide phosphodiesterase (cGI PDE)] isoforms, cGIP1 and cGIP2, have been cloned from rat (R) and human (H) cDNA libraries. The deduced amino acid sequences of RcGIP1 and HcGIP2 are very similar in their conserved catalytic domains but differ in their N-terminal regulatory domains [Meacci, E., et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 3721-3725; Taira, M., et al. (1993) J. Biol. Chem. 268, 18573-18579]. cDNAs encoding both rat adipocyte RcGIP1 and human myocardial HcGIP2 (full-length forms and truncated forms lacking much of the putative N-terminal domain) were expressed in NIH 3006 fibroblasts and in Sf9 insect cells. The recombinant proteins exhibited the expected subunit molecular mass, immunologic reactivities, and characteristics of native membrane-associated forms of the enzymes, e.g., high affinity for cAMP (Km), sensitivity to the selective cGI PDE inhibitors OPC 3689 and OPC 3911 and to cGMP. The full-length recombinants were predominantly particulate, whereas the truncated HcGIP2 forms were cytosolic suggesting that N-terminal domains contain structural determinants important for membrane association. Both fibroblast RcGIP1 and authentic adipocyte cGI PDE were phosphorylated in vitro by cAMP-dependent protein kinase; tryptic [32P]peptides released from rat adipocyte 32P-cGI PDE and 32P-RcGIP1 exhibited identical electrophoretic profiles suggesting that the same peptides are phosphorylated in both.
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- Rahn, Tova, et al.
(författare)
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Identification of the site in the cGMP-inhibited phosphodiesterase phosphorylated in adipocytes in response to insulin and isoproterenol
- 1996
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Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 271:19, s. 11575-11580
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Tidskriftsartikel (refereegranskat)abstract
- Stimulation of rat adipocytes with insulin and isoproterenol results in serine phosphorylation and activation of the adipocyte cGMP-inhibited phosphodiesterase (cGI PDE), events believed to be important in the antilipolytic action of insulin (Degerman, E., Smith, C.J., Tornqvist, H., Vasta, V., Manganiello, V.C., and Belfrage, P. (1990) Proc. Natl. Acad. Sci. U.S.A. 87,533-537). Here we demonstrate, by two-dimensional phosphopeptide mapping, that the major phosphopeptide generated by trypsin, or trypsin followed by Asp-N protease digestion of [32P]cGI PDE phosphorylated in adipocytes in response to isoproterenol and/or insulin, in each case co-migrates with the phosphopeptide released by the same treatment of M297FRRPS(P)LPCISREQ310. This peptide was synthesized based on the deduced sequence of the cloned rat adipocyte cGI PDE and phosphorylated by cAMP-dependent protein kinase (protein kinase A). Radiosequencing of authentic and synthetic tryptic 32P-peptides showed that a single site in cGI PDE (Ser302) was phosphorylated in adipocytes incubated with isoproterenol and/or insulin. The more than additive phosphorylation and activation of cGI PDE in response to the two hormones found in this report and previously (Smith, C.J., Vasta, V., Degerman, E., Belfrage, P., and Manganiello, V.C. (1991) J. Biol. Chem. 266, 13385-13390) is proposed to reflect cross-talk between their respective signal transduction pathways at the level of the cGI PDE serine protein kinase or upstream regulatory component(s).
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- Seidenkrantz, Marit-Solveig, et al.
(författare)
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Two-step deglaciation at the oxygen isotope stage 6/5e transition: the Zeifen-Kattegat climatic oscillation
- 1996
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Ingår i: Quaternary Science Reviews. - 0277-3791 .- 1873-457X. ; 15:1, s. 63-75
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Tidskriftsartikel (refereegranskat)abstract
- Comparison of marine, lacustrine, and terrestrial records from twenty-four sites suggests the existence of a 'Younger Dryas'-type climate oscillation just prior to the Oxygen Isotope Stage 6/5e boundary. These records include results from biostratigraphic, pedostratigraphic, and speleothem studies, as well as analyses of stable isotope compositions of marine records and ice cores. The climate oscillation is named after the warm Zeifen Interstadial and the cold Kattegat Stadial. The Zeifen Interstadial may be related to a major meltwater pulse in the Baffin Bay-Labrador Sea-Norwegian Sea region. The climate oscillation is presumably in part a result of a variation in ocean circulation, especially in the strength of the North Atlantic Drift, but changes in the atmospheric circulation also played an important role. The geographically widespread distribution of the oscillation suggests that the two-step deglaciation influenced both the northern and southern hemispheres.
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