Structural and biochemical characterization of the human cyclophilin family of peptidyl-prolyl isomerases

Autor: Tara L. Davis, Hui Ouyang, Farrell MacKenzie, Sirano Dhe-Paganon, P.J. Finerty, Ragika Paramanathan, John R. Walker, Wolfram Tempel, Elan Z. Eisenmesser, Galina Bernstein, Wen Hwa Lee, Valérie Campagna-Slater
Jazyk: angličtina
Rok vydání: 2010
Předmět:
Gene isoform
Models
Molecular
QH301-705.5
Molecular Sequence Data
Biophysics
Isomerase
Biology
Models
Biological
Biochemistry
General Biochemistry
Genetics and Molecular Biology
Protein Structure
Secondary
03 medical and health sciences
Cyclophilins
Protein structure
Catalytic Domain
Humans
Amino Acid Sequence
Biology (General)
Peptide sequence
Cyclophilin
030304 developmental biology
Peptidylprolyl isomerase
chemistry.chemical_classification
0303 health sciences
General Immunology and Microbiology
General Neuroscience
Biochemistry/Structural Genomics
030302 biochemistry & molecular biology
Biophysics/Structural Genomics
Peptidylprolyl Isomerase
A-site
Enzyme
chemistry
Multigene Family
Biophysics/Biomacromolecule-Ligand Interactions
General Agricultural and Biological Sciences
Research Article
Zdroj: PLoS Biology, Vol 8, Iss 7, p e1000439 (2010)
PLoS Biology
ISSN: 1545-7885
1544-9173
Popis: Peptidyl-prolyl isomerases catalyze the conversion between cis and trans isomers of proline. The cyclophilin family of peptidyl-prolyl isomerases is well known for being the target of the immunosuppressive drug cyclosporin, used to combat organ transplant rejection. There is great interest in both the substrate specificity of these enzymes and the design of isoform-selective ligands for them. However, the dearth of available data for individual family members inhibits attempts to design drug specificity; additionally, in order to define physiological functions for the cyclophilins, definitive isoform characterization is required. In the current study, enzymatic activity was assayed for 15 of the 17 human cyclophilin isomerase domains, and binding to the cyclosporin scaffold was tested. In order to rationalize the observed isoform diversity, the high-resolution crystallographic structures of seven cyclophilin domains were determined. These models, combined with seven previously solved cyclophilin isoforms, provide the basis for a family-wide structure∶function analysis. Detailed structural analysis of the human cyclophilin isomerase explains why cyclophilin activity against short peptides is correlated with an ability to ligate cyclosporin and why certain isoforms are not competent for either activity. In addition, we find that regions of the isomerase domain outside the proline-binding surface impart isoform specificity for both in vivo substrates and drug design. We hypothesize that there is a well-defined molecular surface corresponding to the substrate-binding S2 position that is a site of diversity in the cyclophilin family. Computational simulations of substrate binding in this region support our observations. Our data indicate that unique isoform determinants exist that may be exploited for development of selective ligands and suggest that the currently available small-molecule and peptide-based ligands for this class of enzyme are insufficient for isoform specificity. Enhanced version This article can also be viewed as an enhanced version in which the text of the article is integrated with interactive 3-D representations and animated transitions. Please note that a Web plugin is required to access this enhanced functionality. Instructions for the installation and use of the web plugin are available in Text S1.
Author Summary Cyclophilins are proteins that catalyze the isomerization of prolines, interconverting this structurally important amino acid between cis and trans isomers. Although there are 17 cyclophilins in the human genome, the function of most cyclophilin isoforms is unknown. At least some members of this protein family are of interest for clinically relevant drug design, as they are targets of the drug cyclosporin, which is used as an immunosuppressant to treat patients following organ transplantation. The absence of a comprehensive picture of the similarities and differences between the different members of this protein family precludes effective and specific drug design, however. In the current study we undertake such a global structure∶function analysis. Using biochemical, structural, and computational methods we characterize the human cyclophilin family in detail and suggest that there is a previously overlooked region of these enzymes that contributes significantly to isoform diversity. We propose that this region may represent an important target for isoform-specific drug design.
Databáze: OpenAIRE
Externí odkaz: