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Relationship: 1253

Title

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Binding, Immunophilins leads to Inhibition, Calcineurin Activity

Upstream event
The causing Key Event (KE) in a Key Event Relationship (KER). More help
Downstream event
The responding Key Event (KE) in a Key Event Relationship (KER). More help

Key Event Relationship Overview

The utility of AOPs for regulatory application is defined, to a large extent, by the confidence and precision with which they facilitate extrapolation of data measured at low levels of biological organisation to predicted outcomes at higher levels of organisation and the extent to which they can link biological effect measurements to their specific causes. Within the AOP framework, the predictive relationships that facilitate extrapolation are represented by the KERs. Consequently, the overall WoE for an AOP is a reflection in part, of the level of confidence in the underlying series of KERs it encompasses. Therefore, describing the KERs in an AOP involves assembling and organising the types of information and evidence that defines the scientific basis for inferring the probable change in, or state of, a downstream KE from the known or measured state of an upstream KE. More help

AOPs Referencing Relationship

Taxonomic Applicability

Latin or common names of a species or broader taxonomic grouping (e.g., class, order, family) that help to define the biological applicability domain of the KER.In general, this will be dictated by the more restrictive of the two KEs being linked together by the KER.  More help
Term Scientific Term Evidence Link
Homo sapiens Homo sapiens High NCBI
Mus musculus Mus musculus High NCBI
Rattus norvegicus Rattus norvegicus High NCBI
Macaca mulatta Macaca mulatta High NCBI
Macaca fascicularis Macaca fascicularis High NCBI

Sex Applicability

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Sex Evidence
Mixed High

Life Stage Applicability

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Term Evidence
All life stages High

Key Event Relationship Description

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The phosphatase activity of calcineurin (CN) is well known to be inhibited by CN inhibitors such as FK506 and cyclosporine A through complex formation with immunophilins.

Immunophilins are a general class of proteins that exhibit peptidyl-propyl isomerase (PPIase) activity, such as FKBP (FK506-binding protein) or cyclophilin (Barik. 2006). FKBP and cyclophilin bind with calcineurin (CN)-inhibitors FK506 and cyclosporin A to form complexes, which inhibit CN activity (Barik. 2006).

While FKBP12, FKBP12.6, FKBP13, and FKBP52 are all part of the FK506-binding FKBP family, FKBP12 has a significant involvement in the mechanism of action for FK506-induced immunosuppression (Siekierka et al. 1989, Kang et al. 2008).

FKBP12 is a 12-kDa protein localized in cytoplasm and has been isolated from Jurkat T-cells as a receptor that binds with the CN inhibitor FK506 (Bram et al. 1993). FKBP12 has an FK506-binding domain (FKBD) that comprises 108 amino acids, and is expressed in T‑cells, B‑cells, Langerhans cells, and mast cells (Siekierka et al. 1990, Panhans-Gross et al. 2001, Hultsch et al. 1991).

Cyclophilin and FKBP both exhibit PPIase activity, but no structural similarities have been found between them. Additionally, while immunophilin complexes formed with either substance do inhibit CN phosphatase activity, the PPlase activity and the inhibition of activity that they indicate are unrelated to CN regulation.

CN is a heterodimer that comprises a catalytic subunit (CnA) and a Ca-binding regulatory subunit (CnB). CnA handles phosphatase activity as well as calmodulin binding, and CnB regulates intracellular calcium and CnA (Klee et al. 1988, Zhang et al. 1996). CnA is a 59kDa protein with a serine-threonine phosphatase domain. A FK506-FKBP complex binds directly to CnA in the cell, causing steric hindrance of substrate binding to CN, which in turn inhibits phosphatase activity of CN (Schreiber and Crabtree 1992, Liu et al. 1993, Bierer et al. 1993, Bram et al. 1993, Rao et al. 1997, Liu et al. 1991). Cyclophilin-Cyclosprine A (CsA) complexes also function in the same manner, binding directly to CnA in the cell, which in turn inhibits CN phosphatase activity.

The nuclear factor of activated T cells (NFAT) is a substrate of calcineurin (Rao et al. 1997). When CN activates through stimulus from outside of the cell, it binds directly to the N‑terminal of NFAT in cytoplasm, after which dephosphorylation of SP motifs exposes nuclear localization signal (NLS) and covers nuclear export signal (NES), thereby promoting nuclear localization of NFAT (Matsuda and Koyasu 2000, Zhu and McKeon 1999). When T-cell activation takes place, T-cell receptor (TCR)-mediated stimulus increases the intracellular concentration of calcium and activates CnB, which subsequently induces CnA phosphatase activation, leading to dephosphorylation of NFAT followed by nuclear localization.

When CN activity is inhibited by the binding of immunophilin complexes, dephosphorylation does not occur in NFAT, thereby interfering with nuclear localization.

Evidence Collection Strategy

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Evidence Supporting this KER

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Biological Plausibility
Addresses the biological rationale for a connection between KEupstream and KEdownstream.  This field can also incorporate additional mechanistic details that help inform the relationship between KEs, this is useful when it is not practical/pragmatic to represent these details as separate KEs due to the difficulty or relative infrequency with which it is likely to be measured.   More help

The molecular structures and functions of calcineurin and NFAT are evident based on sufficient scientific findings. The well-known mechanisms for inhibition of calcineurin phosphatase activity by calcineurin inhibitors such as FK506 and cyclosporine A is initiated by their complex formations with their respective immunophilin species. Immunophilins are general classes of proteins that exhibit PPlase activity, but modification of these functions are unrelated to inhibition of CN activity and thought to arise in the molecular structure of the complexes (Schreiber and Crabtree 1992, Liu et al. 1993, Bierer et al. 1993, Bram et al. 1993, Rao et al. 1997, Liu et al. 1991).

It is also well known that inhibition of CN phosphatase activity interfere the dephosphorylation of NFAT leading to suppression of its nuclear localization.

Uncertainties and Inconsistencies
Addresses inconsistencies or uncertainties in the relationship including the identification of experimental details that may explain apparent deviations from the expected patterns of concordance. More help

CN and NFAT are expressed in T cells and other immune cells including B cells, DC and NKT cells, and cytokine productions from these immune cells and expression of IL-2 receptors (IL-2R) in DCs are lowered due to the inhibition of CN phosphatase activity by CN inhibitor treatment. Among them, reduced production of IL-2 and IL-4 from T cells plays a major role in suppression of TDAR as a result of lowed proliferation, differentiation and class switching of B cells, and there have been no reports showing that CN inhibitor-induced reduction of cytokines other than IL-2 and IL-4 as well as reduced expression of IL-2R resulted in TDAR suppression.

FKBP12, a specific immmunophilin that bind with FK506, is also an accessory molecule that bind to IP3 and Ryanodine receptors, both of which are Ca channel located on the membrane of endoplasmic reticulum and participating in the regulation of intracellular Ca concentration.  When binding with FK506, FKBP12 leaves from these receptors to increase the influx of Ca2+ from the endoplasmic reticulum to cytoplasm, which is expected to increase CN activity; however, FK506 treatment suppresses NFAT nuclear localization. In addition, FKBP12-knock out mice show no changes in immune functions including T cell functions.  These facts suggest that inhibition of CN-NFAT system induced by FK506 treatment result from direct inhibition of CN phosphatase activity by FK506-FKBP12 complex and not by affecting Ryanodine and IP3 receptors associated with FKBP12.

Known modulating factors

This table captures specific information on the MF, its properties, how it affects the KER and respective references.1.) What is the modulating factor? Name the factor for which solid evidence exists that it influences this KER. Examples: age, sex, genotype, diet 2.) Details of this modulating factor. Specify which features of this MF are relevant for this KER. Examples: a specific age range or a specific biological age (defined by...); a specific gene mutation or variant, a specific nutrient (deficit or surplus); a sex-specific homone; a certain threshold value (e.g. serum levels of a chemical above...) 3.) Description of how this modulating factor affects this KER. Describe the provable modification of the KER (also quantitatively, if known). Examples: increase or decrease of the magnitude of effect (by a factor of...); change of the time-course of the effect (onset delay by...); alteration of the probability of the effect; increase or decrease of the sensitivity of the downstream effect (by a factor of...) 4.) Provision of supporting scientific evidence for an effect of this MF on this KER. Give a list of references.  More help
Response-response Relationship
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Time-scale
Information regarding the approximate time-scale of the changes in KEdownstream relative to changes in KEupstream (i.e., do effects on KEdownstream lag those on KEupstream by seconds, minutes, hours, or days?). More help
Known Feedforward/Feedback loops influencing this KER
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Domain of Applicability

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FKBP is found in a wide variety of organisms, from prokaryotes to multicellular organisms (Siekierka et al. 1989). Multiple subfamilies of FKBP have been reported, with at least eight types having been found in mammals. FKBP12 is reported to be expressed in B-cells, Langerhans cells, and mast cells as well as in T-cells of humans, mice, and other mammalian species.

Cyclophilins have been found in mammals, plants, insects, fungi, and bacteria. They are structurally conserved throughout evolution and all have PPIase activity (Wang P et al. 2005).

CN is broadly distributed throughout the body, including T- and B-cells, and the structure of CnA and CnB is highly conserved from yeasts to humans. Also highly conserved are the amino acid sequences of the catalytic and regulatory domains of CnA isoforms from different organisms (Kincaid. 1993).

NFAT expresses in B cells, mast cells, neutrophil granulocytes, dendritic cells, macrophages, and natural killer cells as well as T cells from humans, rodents, and other mammalian species (Rao et al. 1997).

References

List of the literature that was cited for this KER description. More help
  1. Barik, S. (2006). Immunophilins: for the love of proteins. Cellular and Molecular Life Sciences 63(24): 2889-900.
  2. Bierer, B.E., Holländer, G., Fruman, D. and Burakoff, S.J. (1993). Cyclosporin A and FK506: molecular mechanisms of immunosuppression and probes for transplantation biology. Current opinion in immunology 5 (5): 763-73.
  3. Bram, R.J., Hung, D.T., Martin, P.K., Schreiber, S.L. and Crabtree, G.R. (1993). Identification of the immunophilins capable of mediating inhibition of signal transduction by cyclosporin A and FK506: roles of calcimeurin binding and cellular location. Molecular and cellular biology 13 (8): 4760-9.
  4. Flanagan, W.M., Corthésy, B., Bram, R.J. and Crabtree, G.R. (1991). Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A. Nature 352 (6338): 803-7.
  5. Fruman, D. A., Klee, C. B., Bierer, B. E. and Burakoff, S. J. (1992). Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A. Proceedings of the National Academy of Sciences of the United States of America. 89(9):3686-90.
  6. Fruman, D. A., Bierer, B. E., Benes, J. E., Burakoff, S. J., Austen, K. F. and Katz, H. R. (1995). The complex of FK506-binding protein 12 and FK506 inhibits calcineurin phosphatase activity and IgE activation-induced cytokine transcripts, but not exocytosis, in mouse mast cells. Journal of Immunology.154(4):1846-51.
  7. Hultsch, T., Albers, M. W., Schreiber, S.L. and Hohman, R. J. (1991). Immunophilin ligands demonstrate common features of signal transduction leading to exocytosis or transcription. Proceedings of the national academic science of the United States of America. 14: 6229-6233.
  8. Kang, C. B., Hong, Y., Dhe-Paganon, S. and Yoon, H. S. (2008). FKBP family proteins: immunophilins with versatile biological functions. Neurosignals. 16: 318-325.
  9. Kincaid, R .L. (1993). Calmodulin-dependent protein phosphatases from microorganisms to man. A study in structural conservatism and biological diversity. Adv Second Messenger Phosphoprotein Res. 27:1-23.
  10. Klee, C. B., Draetta, G. F. and Hubbard, M. J. (1988). Calcineurin. Advances in enzymology and related areas of molecular biology. 61:149-200.
  11. Liu, J., Farmer, J. D. Jr., Lane, W. S., Friedman, J., Weissman, I. and Schreiber, S. L. (1991). Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell. 66(4): 807-815.
  12. Liu, J., Albers, M. W., Wandless, T. J., Luan, S., Alberg, D. G., Belshaw, P. J., Cohen, P., MacKintosh, C., Klee, C. B. and Schreiber, S.L.. (1992). Inhibition of T cell signaling by immunophilin-ligand complexes correlates with loss of calcineurin phosphatase activity. Biochemistry. 31(16):3896-901.
  13. Liu, J. (1993). FK506 and cyclosporin, molecular probes for studying intracellular signal transduction. Immunology today. 14(6): 290-305.
  14. Maguire O, Tornatore KM, O'Loughlin KL, Venuto RC and Minderman H. (2013) Nuclear translocation of nuclear factor of activated T cells (NFAT) as a quantitative pharmacodynamic parameter for tacrolimus. Cytometry A. 83(12):1096-104.
  15. Matsuda, S., Koyasu, S. (2000). A second target of cyclosporin A and FK506. Tanpakushitsu kakusan koso. 45(11): 1823-1831.
  16. Panhans-Gross, A., Novak, N., Kraft, S. and Bieber, T. (2001). Human epidermal Langerhans' cells are targets for the immunosuppressive macrolide tacrolimus (FK506). Journal of Allergy and Clinical Immunology 107(2): 345-52.
  17. Rao, A., Luo, C. and Hogan, PG. (1997). Transcription factors of the NFAT family: regulation and function. Annual Review of Immunology 15: 707-47.
  18. Schreiber, SL. and Crabtree, GR. (1992). The mechanism of action of cyclosporin A and FK506. Immunology Today 13(4): 136-42. >
  19. Siekierka, JJ., Hung, SH., Poe, M., Lin, CS. and Sigal, NH. (1989). A cytosolic binding protein for the immunosuppressant FK506 has peptidyl-prolyl isomerase activity but is distinct from cyclophilin. Nature 341(6244): 755-57.
  20. Siekierka, JJ., Wiederrecht, G., Greulich, H., Boulton, D., Hung, SH., Cryan, J., Hodges, PJ. and Sigal, NH. (1990). The cytosolic-binding protein for the immunosuppressant FK-506 is both a ubiquitous and highly conserved peptidyl-prolyl cis-trans isomerase. Journal of Biological Chemistry 265(34): 21011-5.
  21. Wang, P. and Heitman, J. (2005) The cyclophilins. Genome Biology 6 (7):226.
  22. Zhang, B.W., Zimmer, G., Chen, J., Ladd, D., Li, E., Alt, F.W., Wiederrecht, G., Cryan, J., O'Neill, E.A., Seidman, C.E., Abbas, A.K. and Seidman, J.G. (1996). T cell responses in calcineurin A alpha-deficient mice. Journal of experimental medicine 183(2): 413-20.
  23. Zhu, J. and McKeon, F. (1999). NF-AT activation requires suppression of Crm1-dependent export by calcineurin. Nature. 398(6724): 256-60.