Further evidence for abnormal protein kinase C regulation of macromolecule secretion in fibroblasts from cystic fibrosis patients |
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Authors: | F Bertrand B Hermelin A Paul I Garcia J Capeau G Cherqui J Picard |
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Institution: | (1) Laboratoire de Biochimie, Hôpital Sainte-Eugénie, 69310 Pierre Bénite, France;(2) Laboratoire de Biochimie-Biologie Cellulaire, I.N.S.E.R.M.-U. 181, Faculté de Médecine Saint-Antoine, 27, rue Chaligny, 75571 Paris Cédex 12, France |
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Abstract: | In comparison to skin fibroblasts from normal subjects, those from patients with cystic fibrosis (CF): (1) bound 20-3H] phorbol 12,13-dibutyrate (PDBu) with a higher affinity (Kd=25.8 vs 12.8 nM respectively) but expressed a similar number of total phorbol ester binding sites (about 2.5 pmol PDBu bound/mg of protein); (2) exhibited a faster and higher response to 4 -phorbol 12 -myristate 13 -acetate (PMA) for the stimulation of 35S]-labelled glycoconjutate release, but were equally sensitive to the synergistic effect of A23187 on this process; and (3) secreted glycoconjugates with similar 35S]-sulfate and 14C]-leucine to 14C]-glucosamine labelling ratios. Taken together, these results provide further evidence for abnormal protein kinase C (PKC) regulation of macromolecule secretion in CF disease.Abbreviations BSA
Bovine serum albumin
- DBcAMP
Dibutyryl cyclic AMP
- DMEM
Dulbecco's modified Eagle's medium
- DMSO
Dimethylsulfoxide
- LDH
Lactate dehydrogenase
- PBS
Phosphate-buffered saline
- PDBu
4 -phorbol 12,13-dibutyrate
- 4 -PDD
4 -phorbol 12,13-didecanoate
- PMA
4 -phorbol 12 -myristate 13 -acetate
- TCA
Trichloroacetic acid |
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Keywords: | cystic fibrosis protein kinase C glycoconjugate secretion phorbol esters |
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