Differential expression of cyclophilin isoforms during keratinocyte differentiation.
Cyclophilin A, the major intracellular binding protein for the immunosuppressive drug cyclosporin A (CsA), was studied in human keratinocytes during differentiation both in vivo and in vitro. Analysis of cyclophilin by gel-filtration radiobinding-assay with tritiated CsA showed one specific radioactive peak at 17 kDa. By this technique, the levels of cyclophilin (mean 55.23 +/- 8.43 pmol/mg protein) did not significantly differ during keratinocyte differentiation. When the protein extracts from calcium-induced differentiating keratinocytes and normal human skin were analysed by PAGE radiobinding-assay, two specific radioactive CsA-binding peaks were detected. The major peak (RF 0.13) was expressed in all samples (mean 47.32 +/- 17.53 pmol/mg protein) whereas the minor peak (RF 0.23) was dramatically decreased about 6-fold in abnormally differentiated skin (psoriasis) as well as in non-differentiated keratinocytes. At least six [3H]CsA-binding isoforms with pI values ranging from 5.58 to 7.75 were detected by isoelectrofocusing autoradio-blotting-assay in normal human skin; three of them immunoreacted with antibodies to cyclophilin. These results demonstrated the presence of several cyclophilin isoforms in human epidermal cells and an expression which correlated with the differentiation of human keratinocytes both in vivo and in vitro.
Chatellard-Gruaz
D
D
Clinique de Dermatologie, Hôpital Cantonal Universitaire, Genève, Switzerland.
Saurat
J H
JH
Siegenthaler
G
G
D000591:Amino Acid Isomerases / Q000302:isolation & purification / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007525:Isoelectric Focusing; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism*; D019696:Peptidylprolyl Isomerase
D002352:Carrier Proteins; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase