Identification of low abundance cyclophilins in human plasma. Issue 21 (12th October 2016)

• Record Type: Journal Article
• Authors: Schumann, Michael; Ihling, Christian H.; Prell, Erik; Schierhorn, Angelika; Sinz, Andrea; Fischer, Gunter; Schiene‐Fischer, Cordelia; Malešević, Miroslav
• Is Part Of: Proteomics. Volume 16:Issue 21(2016)
• Page Start: 2815
• Page End: 2826
• Publication Date: 2016-10-12
• Subjects: Acetylation -- Affinity‐proteomics -- Biomedicine -- Cyclophilin -- Cyclosporine A -- PPIase; Proteins -- Separation -- Periodicals; Bioinformatics -- Periodicals; Proteomics -- Periodicals; Genomes -- Periodicals; Molecular genetics -- Periodicals; 572.605
• DOI: 10.1002/pmic.201600221
• Languages: English
• ISSNs: 1615-9853

Abstract

Abstract : Cylophilins (Cyps) belong to the ubiquitously distributed enzyme class of peptidyl prolyl cis/trans isomerases (EC5.2.1.8), which are foldases capable of accelerating slow steps in the refolding of denatured proteins. At least 20 different Cyp isoenzymes are broadly distributed among all organs and cellular compartments in humans. Extracellularly localized Cyps came into the scientific focus recently because of their involvement in the control of inflammatory diseases, as well as viral and bacterial infections. However, detailed insights into Cyp functions are often hampered by the lack of sensitive detection methods. We present an improved method for affinity purification and detection of Cyp in biotic samples in this manuscript. The procedure takes advantage of two novel cyclosporine A derivatives. Derivative1 was used to capture Cyps from the sample while derivative2 was applied for selective release from the affinity matrix. Using this approach, eight different Cyp (CypA, CypB, CypC, Cyp40 (PPID), CypE, CypD (PPIF), CypH, and CypL1) were unambiguously detected in healthy human blood plasma. Moreover, extracellular CypA was found to be partially modified by N ε acetylation on residues Lys44, Lys133, Lys155, as well as N α acetylation at the N ‐terminal Val residue. N α acetylation of Ser2 residue was also found for Cyp40.