title: Biochemical Features of Recombinant Human Cyclophilin J.
abstract: OBJECTIVE
  To characterize the biochemical features of the newest member of cyclophilin family of peptidyl-prolyl cis/trans-isomerases (PPIases), cyclophilin J (CYPJ).
  
  
  METHODS
  PPIase assays were performed on purified hCYPJ and its mutated variants. The substrate specificity, half-maximal inhibitory concentration (IC50) of cyclosporin A (CsA) inhibition and circular dichroism (CD) spectrum of CYPJ were measured. Mercury pathway profiling luciferase assays were also performed.
  
  
  RESULTS
  The catalytic number/Michaelis constant (kcat/KM) value of CYPJ was 9.5×10(4) s(-1)M(-1). CYPJ additionally catalyzed norleucine-proline, isoleucine-proline and glutamine-proline peptides compared to CYPA and Escherichia coli PPIases. CYPJ was inhibited by CsA in a dose-dependent manner with IC50 of 12.1±0.9 μM. The CD spectrum of CYPJ was similar to CYPA. CYPJ significantly up-regulated the transcription of E-box, E2F, retinoblastoma (Rb), p53, activator protein 1 (AP1), NF-κB and phospho-cAMP response element (CRE) cis-response element in 293T cells.
  
  
  CONCLUSIONS
  CYPJ structurally resembles CYPA. It is sensitive to inhibition by CsA and plays a role in regulating cell growth, proliferation, and apoptosis.
authors.affiliation: Department of Oncology, Yantai Yuhuangding Hospital, Affiliated Hospital of Qingdao University, Yantai, Shandong, P.R. China State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, P.R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.affiliation: MPG-Research-Unit Enzymology of Protein Folding, Halle/Saale, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Liefke
authors.affiliation: Department of Oncology, Yantai Yuhuangding Hospital, Affiliated Hospital of Qingdao University, Yantai, Shandong, P.R. China.
authors.forename: Lixin
authors.initials: L
authors.lastname: Jiang
authors.affiliation: Department of Oncology, Yantai Yuhuangding Hospital, Affiliated Hospital of Qingdao University, Yantai, Shandong, P.R. China.
authors.forename: Jiahui
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, P.R. China.
authors.forename: Chaoqun
authors.initials: C
authors.lastname: Huang
authors.affiliation: Department of Oncology, Yantai Yuhuangding Hospital, Affiliated Hospital of Qingdao University, Yantai, Shandong, P.R. China State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, P.R. China gongzhaohuayt@163.com schiene@enzyme-halle.mpg.de.
authors.forename: Zhaohua
authors.initials: Z
authors.lastname: Gong
authors.affiliation: MPG-Research-Unit Enzymology of Protein Folding, Halle/Saale, Germany gongzhaohuayt@163.com schiene@enzyme-halle.mpg.de.
authors.forename: Cordelia
authors.initials: C
authors.lastname: Schiene-Fischer
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, P.R. China.
authors.forename: Long
authors.initials: L
authors.lastname: Yu
mesh_terms: D002384:Catalysis; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D021983:Cyclophilins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D016572:Cyclosporine / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D050684:E2F Transcription Factors / Q000235:genetics / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D009154:Mutation; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: C115163:CREB1 protein, human; D017362:Cyclic AMP Response Element-Binding Protein; D050684:E2F Transcription Factors; D004791:Enzyme Inhibitors; D016328:NF-kappa B; D011994:Recombinant Proteins; D016160:Retinoblastoma Protein; C495901:TP53 protein, human; D018808:Transcription Factor AP-1; D016159:Tumor Suppressor Protein p53; D016572:Cyclosporine; D021983:Cyclophilins; C503209:PPIL3 protein, human
keywords: CYPJ; Cyclophilin J; PPIL3; PPIase activity; biochemistry