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Multiple pertusis toxin substrates as candidates for regulatory G proteins of adenylate cyclase coupled to the somatostatin receptor in primary rat astrocytes

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Abstract

The involvement of G proteins in receptor mediated astroglial cAMP formation was studied. Isoproterenol or prostaglandin E2 stimulated adenylate cyclase of primary astroglial cells was inhibited by somatostatin. Preincubation, of cells with increasing concentrations of islet activating protein (IAP) diminished somatostatin inhibition of adenylate cyclase. At an IAP concentration of 50 ng/ml somatostatin inhibition was completely abolished. Studies on IAP catalyzed32P-ADP-ribosylation of astroglial cell particulate material revealed an incorporation of radiolabel into three polypeptides in the molecular weight range of 41,000–39,000 Dalton. Pretreatment of intact cells with IAP reduced radiolabeling of this molecular species in a concentration dependent manner. No further radiolabeling above background level was detectable after pretreatment of cultures with 10 ng IAP/ml or more. At present, the occurrence of at least three IAP substrates (G proteins) does not permit an identification of the somatostatin receptor coupled G protein. Rather, the finding reveals that astrocytes are endowed with multiple variants of GTP binding proteins likely to be coupled to different receptors.

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Abbreviations

cAMP:

cyclic adenosine monophosphate

DPBS:

Dulbecco's phosphate buffered saline

DTT:

dithiothreitol

IAP:

islet activating protein=pertussis toxin

PG:

prostaglandin

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Authors and Affiliations

  1. Pharmacological Institute, University of Freiburg, Hermann-Herder-Str. 5, D-7800, Freiburg i.Br., F.R.G.

    Peter J. Gebicke-Haerter, Siegfried Wurster, Angelika Schobert, Clemens Allgaier & Georg Hertting

  2. Institute of Experimental Medicine, Hungarian Academy of Sciences, P.O. Box 67, H-1450, Budapest, Hungary

    András Seregi

Authors
  1. Peter J. Gebicke-Haerter
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  2. András Seregi
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  3. Siegfried Wurster
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  4. Angelika Schobert
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  5. Clemens Allgaier
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  6. Georg Hertting
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Gebicke-Haerter, P.J., Seregi, A., Wurster, S. et al. Multiple pertusis toxin substrates as candidates for regulatory G proteins of adenylate cyclase coupled to the somatostatin receptor in primary rat astrocytes. Neurochem Res 13, 997–1001 (1988). https://doi.org/10.1007/BF00970774

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