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Inhibitory guanine nucleotide binding protein in pig epidermis: regulation of epidermal adenylate cyclase

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Summary

Islet-activating protein (IAP), one of the pertussis toxins, serving [α-32P]nicotinamide adenine dinucleotide (NAD) as a substrate for ADP ribosylation, radiolabelled a specific pig epidermal membrane protein. The IAP-specific substrate was detectable by sodium dodecyl sulphate-polyacrylamide gel electrophoresis as a single band corresponding to a molecular weight of 40 kDa. The ADP ribosylation catalysed by IAP was inhibited by the addition of Mg2+ to the reaction mixture. IAP is known to work on intact cell systems resulting in the ADP ribosylation using intracellular NAD as the ADP ribose donor. Following IAP pretreatment of intact pig epidermis, the epidermal receptor adenylate cyclase responses were markedly increased; all the stimulatory receptor adenylate cyclase reponses (beta-adrenergic, prostaglandin E, adenosine and histamine responses) were significantly increased. Cholera toxin-induced cyclic AMP accumulation was also significantly increased. Forskolin-induced cyclic AMP accumulation was slightly increased after IAP pretreatment, but this was not statistically significant. The IAP-dependent ADP ribosylation of the epidermal 40 kDa membrane protein, which was prepared from the IAP pretreated epidermis, was significantly decreased. It is known that the tumour promoter, phorbol 12-myristate,13-acetate (PMA), decreases stimulatory receptor adenylate cyclase responses of the epidermis. Following the PMA pretreatment, IAP-dependent ADP ribosylation of the epidermal membrane protein was unaffected. Furthermore, following the PMA pretreatment, the IAP-induced increase in the epidermal receptor adenylate cyclase responses still remained. Our results indicate that pig epidermis contains 40 kDa membrane substrate for IAP-dependent ADP ribosylation, which has an inhibitory tonus on the epidermal adenylate cyclase until its ADP ribosylation by IAP. The results are consistent with the view that the epidermis contains an IAP-sensitive inhibitory guanine nucleotide binding protein (Gi) of the adenylate cyclase system. Although PMA decreases epidermal stimulatory receptor adenylate cyclase responses, it is unlikely that PMA reveals its effect through the modulation of Gi.

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Authors and Affiliations

  1. Department of Dermatology, Asahikawa Medical College, 3-11 Nishikagura Asahikawa, 078, Japan, Japan

    M. Tsutsui & H. Iizuka

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  1. M. Tsutsui
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  2. H. Iizuka
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Tsutsui, M., Iizuka, H. Inhibitory guanine nucleotide binding protein in pig epidermis: regulation of epidermal adenylate cyclase. Arch Dermatol Res 282, 469–474 (1990). https://doi.org/10.1007/BF00402625

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  • DOI: https://doi.org/10.1007/BF00402625

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