title: Identification of low abundance cyclophilins in human plasma.
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. Derivative 1 was used to capture Cyps from the sample while derivative 2 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 Nεacetylation on residues Lys44, Lys133, Lys155, as well as Nαacetylation at the N-terminal Val residue. Nαacetylation of Ser2 residue was also found for Cyp40.
authors: Department of Enzymology, Institute of Biochemistry und Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle, Germany.
authors: Michael
authors: M
authors: Schumann
authors: Department of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin-Luther-University Halle-Wittenberg, Halle, Germany.
authors: Christian H
authors: CH
authors: Ihling
authors: Branch Office Halle, Max-Planck Institute for Biophysical Chemistry, Göttingen, Halle, Germany.
authors: Erik
authors: E
authors: Prell
authors: Department of Enzymology, Institute of Biochemistry und Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle, Germany.
authors: Angelika
authors: A
authors: Schierhorn
authors: Department of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin-Luther-University Halle-Wittenberg, Halle, Germany.
authors: Andrea
authors: A
authors: Sinz
authors: Branch Office Halle, Max-Planck Institute for Biophysical Chemistry, Göttingen, Halle, Germany.
authors: Gunter
authors: G
authors: Fischer
authors: Department of Enzymology, Institute of Biochemistry und Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle, Germany.
authors: Cordelia
authors: C
authors: Schiene-Fischer
authors: Department of Enzymology, Institute of Biochemistry und Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle, Germany. miroslav.malesevic@biochemtech.uni-halle.de.
authors: Miroslav
authors: M
authors: Malešević
mesh_terms: D000107:Acetylation; D002851:Chromatography, High Pressure Liquid; D021983:Cyclophilins / Q000097:blood* / Q000145:classification; D016572:Cyclosporine / Q000097:blood* / Q000145:classification; D006801:Humans; D020543:Proteome / Q000235:genetics*; D040901:Proteomics
chemical_list: D020543:Proteome; D016572:Cyclosporine; D021983:Cyclophilins
keywords: Acetylation; Affinity-proteomics; Biomedicine; Cyclophilin; Cyclosporine A; PPIase