Biochemical Features of Recombinant Human Cyclophilin J

Jian Chen; Robert Liefke; Lixin Jiang; Jiahui Wang; Chaoqun Huang; Zhaohua Gong; Cordelia Schiene-Fischer; Long Yu

Biochemical Features of Recombinant Human Cyclophilin J

Anticancer Res. 2016 Mar;36(3):1175-80.

Authors

Jian Chen¹, Robert Liefke², Lixin Jiang³, Jiahui Wang³, Chaoqun Huang⁴, Zhaohua Gong⁵, Cordelia Schiene-Fischer⁶, Long Yu⁴

Affiliations

¹ 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.
² MPG-Research-Unit Enzymology of Protein Folding, Halle/Saale, Germany.
³ Department of Oncology, Yantai Yuhuangding Hospital, Affiliated Hospital of Qingdao University, Yantai, Shandong, P.R. China.
⁴ State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, P.R. China.
⁵ 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 [email protected] [email protected].
⁶ MPG-Research-Unit Enzymology of Protein Folding, Halle/Saale, Germany [email protected] [email protected].

PMID: 26977013

Abstract

Aim: To characterize the biochemical features of the newest member of cyclophilin family of peptidyl-prolyl cis/trans-isomerases (PPIases), cyclophilin J (CYPJ).

Materials and 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.

Conclusion: CYPJ structurally resembles CYPA. It is sensitive to inhibition by CsA and plays a role in regulating cell growth, proliferation, and apoptosis.

Keywords: CYPJ; Cyclophilin J; PPIL3; PPIase activity; biochemistry.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Catalysis
Cyclic AMP Response Element-Binding Protein / genetics
Cyclic AMP Response Element-Binding Protein / metabolism
Cyclophilins / antagonists & inhibitors
Cyclophilins / genetics
Cyclophilins / metabolism*
Cyclosporine / metabolism
Cyclosporine / pharmacology
Dose-Response Relationship, Drug
E2F Transcription Factors / genetics
E2F Transcription Factors / metabolism
Enzyme Inhibitors / metabolism
Enzyme Inhibitors / pharmacology
Gene Expression Regulation
HEK293 Cells
Humans
Kinetics
Mutation
NF-kappa B / genetics
NF-kappa B / metabolism
Protein Binding
Recombinant Proteins / metabolism
Retinoblastoma Protein / genetics
Retinoblastoma Protein / metabolism
Substrate Specificity
Transcription Factor AP-1 / genetics
Transcription Factor AP-1 / metabolism
Transcription, Genetic
Transfection
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism

Substances

CREB1 protein, human
Cyclic AMP Response Element-Binding Protein
E2F Transcription Factors
Enzyme Inhibitors
NF-kappa B
Recombinant Proteins
Retinoblastoma Protein
TP53 protein, human
Transcription Factor AP-1
Tumor Suppressor Protein p53
Cyclosporine
Cyclophilins
PPIL3 protein, human