Keratinocyte Secretion of Cyclophilin B via the Constitutive Pathway Is Regulated through Its Cyclosporin-Binding Site

• Published in: Journal of investigative dermatology Vol. 131; no. 5; pp. 1085 - 1094
• Main Authors: Fearon, Paula, Lonsdale-Eccles, Ann A., Ross, O. Kehinde, Todd, Carole, Sinha, Aparna, Allain, Fabrice, Reynolds, Nick J.
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.05.2011; Nature Publishing Group; Elsevier Limited
• Subjects: Binding Sites; Biological and medical sciences; Cells, Cultured; Cyclophilins - metabolism; Cyclosporine - pharmacology; Dermatologic Agents - pharmacology; Dermatology; Endoplasmic Reticulum - drug effects; Endoplasmic Reticulum - enzymology; Golgi Apparatus - drug effects; Golgi Apparatus - enzymology; Humans; Keratinocytes - drug effects; Keratinocytes - enzymology; Medical sciences; Original; Secretory Vesicles - drug effects; Secretory Vesicles - enzymology; Regulation(control); Calcineurin inhibitor; Dermatology; Secretion; Keratinocyte; Ciclosporin; Binding site; Immunosuppressive agent
• ISSN: 0022-202X; 1523-1747
• DOI: 10.1038/jid.2010.415

Cyclophilin B (CypB) is an endoplasmic reticulum (ER)-resident member of the cyclophilin family of proteins that bind cyclosporin A (CsA). We report that as in other cell types, CypB trafficked from the ER and was secreted by keratinocytes into the media in response to CsA. Concentrations as low as 1pM of CsA induced secretion of CypB. Using brefeldin A, we showed that CypB is secreted from keratinocytes via the constitutive secretory pathway. We defined that substitution of tryptophan residue 128 in the CsA-binding site of CypB with alanine resulted in dissociation of CypBW128A-green fluorescent protein (GFP) from the ER. Photobleaching studies revealed a significant reduction in the diffusible mobility of CypBW128A-GFP compared with CypBWT-GFP, consistent with redistribution of CypBW128A-GFP into secretory vesicles disconnected from the ER/Golgi network. Furthermore, CsA significantly decreased the mobility of CypBWT-GFP but not CypBW128A-GFP. These studies demonstrate that therapeutically relevant concentrations of CsA regulate secretion of CypB by keratinocytes, and that a key residue within the CsA-binding site of CypB controls retention of CypB within the ER and regulates entry into the secretory pathway. As keratinocytes express CypB receptors (CD147) and CypB exhibits chemotactic properties, these data have implications for the therapeutic effects of CsA in inflammatory skin disease.