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  A system to select for mutant LLC-PK1 cells affected in cAMP mediated hormonal response using a photoactivatable analogue of vasopressin

Luzius, H., Jans, D. A., & Fahrenholz, F. (1990). A system to select for mutant LLC-PK1 cells affected in cAMP mediated hormonal response using a photoactivatable analogue of vasopressin. Journal of Receptors and Signal Transduction, 10(1-2), 61-80. doi:10.3109/10799899009064658.

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 Creators:
Luzius, Heike1, Author              
Jans, David A.1, Author              
Fahrenholz, Falk1, Author              
Affiliations:
1Emeritusgroup Physical Chemistry, Max Planck Institute of Biophysics, Max Planck Society, ou_3273414              

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 Abstract: The photoreactive analogue of vasopressin, [1-(3-mercapto)propionic acid, 8-(N6-4-azidophenyl-amidino)lysine] vasopressin (apa-LVP) could be used to elicit stimulation of cAMP production in LLC-PK renal epithelial cells, detectable up to 24 h after photoactivation by flash photolysis. This is in contrast to cells treated with vasopressin, or apa-LVP without photoactivation, where cAMP synthesis is down regulated within 4 h. The prolonged stimulation of cAMP production induced by photoactivation of apa-LVP was demonstrated to be cytotoxic to LLC-PK1 cells, whereas the vasopressin receptor negative LLC-PK1 mutant M18 was resistant to the cytotoxic effect. A selection strategy was developed for mutants resistant to this long-term stimulation of cAMP production, whereby multiple cycles of treatment with apa-LVP and photoactivation were used. Mutants so selected were then characterized using a novel screening system for detection of the production of urokinase-type plasminogen activator in response to cAMP agonists. One mutant was examined and found to be impaired in hormonal responsiveness, whereby hormone and forskolin stimulated cAMP-mediated responses were markedly reduced. It exhibited resistance to the long-term stimulation of cAMP production elicited by apa-LVP and photoactivation. This implies that apa-LVP can be used to select for novel mutants specifically impaired in cAMP metabolism and in particular down-regulation of cAMP response.

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Language(s): eng - English
 Dates: 2008-09-261990
 Publication Status: Published in print
 Pages: 20
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.3109/10799899009064658
PMID: 2175811
 Degree: -

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Title: Journal of Receptors and Signal Transduction
Source Genre: Journal
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Publ. Info: New York, NY : Marcel Dekker
Pages: - Volume / Issue: 10 (1-2) Sequence Number: - Start / End Page: 61 - 80 Identifier: ISSN: 1079-9893
CoNE: https://pure.mpg.de/cone/journals/resource/954927700049