Cyclosporine—relationship of Side Effects to Mode of Action

Ryffel, Bernhard; Foxwell, Brian M. J.; Gee, Andrew; Greiner, Brigitte; Woerly, G.; Mihatsch, Michael J.

doi:10.1097/00007890-198808001-00017

Cited by 59 publications

(25 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…CSA is a known ligand for cyclophilin D, a mitochondrial matrix protein belonging to the peptidyl-prolyl cis,transisomerase family that is associated with mitochondrial voltagedependent ion channels. CSA prevents opening of the PTPs and release of cytochrome c into the cytoplasm in response to binding of the proapoptotic proteins Bax and Bak (12,16,42,52). Perhaps the ability of CSA to protect mitochondria is not limited to inhibition of the opening of PTPs but instead represents a general strengthening of the physical properties of the MOM.…”

Section: Discussionmentioning

confidence: 99%

Dynamin-2-Dependent Targeting ofMannheimia haemolyticaLeukotoxin to Mitochondrial Cyclophilin D in Bovine Lymphoblastoid Cells

et al. 2008

View full text Add to dashboard Cite

Exotoxins which belong to the family containing the RTX toxins (repeats in toxin) contribute to a variety of important human and animal diseases. One example of such a toxin is the potent leukotoxin (LKT) produced by the bovine respiratory pathogen Mannheimia haemolytica. LKT binds to CD18, resulting in the death of bovine leukocytes. In this study, we showed that internalized LKT binds to the outer mitochondrial membrane, which results in the release of cytochrome c and collapse of the mitochondrial membrane potential ( M. haemolytica is the principal bacterial pathogen in the bovine respiratory disease complex (19, 58). The most important virulence factor of M. haemolytica is the LKT, which has potent cytotoxic effects on ruminant leukocytes but not on leukocytes from other species (38). M. haemolytica LKT is a member of the RTX toxin (repeats in toxin) family, which includes LKTs and hemolysins produced by a number of gramnegative bacteria. It was originally thought that M. haemolytica LKT damages cells by inserting into the cell membrane, resulting in pore formation and necrosis (53). At lower LKT concentrations, however, susceptible cells die via caspase-dependent apoptotic pathways (8,10,45,46). LKT was first reported to bind to the ␤ 2 integrin CD18/CD11a, also known as leukocyte functional antigen 1 (LFA-1) (1,24,28). Recent studies, however, have shown that Mac-1 (CD18/CD11b) also binds LKT and that CD18 is the functional receptor for LKT (9, 30). After binding to LFA-1, LKT induces apoptosis of bovine leukocytes as a result of activation of caspase 1, caspase 3, and caspase 9 (4, 10, 31, 46). The LFA-1-dependent cytotoxicity of LKT is a conundrum, because signaling through LFA-1 generally results in cell adhesion and promotes cell survival (34,44,51).In this study we hypothesized that mitochondrial damage and caspase 9 activation could be due to direct binding of LKT to the mitochondrial outer membrane (MOM). In this paper we show that LKT directly targets mitochondria in BL-3 cells, creating lesions in the MOM that can lead to release of proapoptotic proteins, culminating in cell death. We also show that LKT targeting to mitochondria is dynamin-2 dependent and that mitochondria appear to be the principal source of dynamin-2 in BL-3 cells. Treating BL-3 cells with cyclosporine (CSA) depleted mitochondrial dynamin-2, thereby inhibiting LKT transport to mitochondria and cell death. MATERIALS AND METHODSLKT production and purification. Crude LKT was prepared and purified as described previously and was stored at Ϫ80°C until it was used in experiments (4). Inactive toxin from an lktC mutant of M. haemolytica strain A1 (SH 1562), which produces an LKT protein with no biological activity (LKT inact ) (generously provided by S. K. Highlander, Baylor College of Medicine, Houston, TX), was prepared in a similar manner.Cell lines, cell cultures, and antibodies. Nonadherent bovine lymphoblastoid cells (BL-3 cells) and adherent murine macrophages (RAW 264.7 cells) (kindly provided by Ronald Schultz, University of...

show abstract

Section: Discussionmentioning

confidence: 99%

Dynamin-2-Dependent Targeting ofMannheimia haemolyticaLeukotoxin to Mitochondrial Cyclophilin D in Bovine Lymphoblastoid Cells

et al. 2008

View full text Add to dashboard Cite

show abstract

“…To completely convert chromatin, which contains 6 ϫ 10 9 nucleotides (59), to 50-kb fragments, 60,000 phosphodiester bonds must be cleaved. Human thymocytes contain approximately 540,000 molecules of CypA (based on an average of 1.6 g of CypA/mg of cell protein) (8). Therefore, only 60,000, or 11%, of all CypA molecules in the thymocyte need to be in contact with the chromatin and to make one disruption of the phosphodiester bond to completely reduce the chromatin to 50-kb fragments.…”

Section: Discussionmentioning

confidence: 99%

“…This large family of proteins has been isolated from a diverse range of organisms, including Neurospora (4), Escherichia coli (5,6), yeast (7), plants, and higher eukaryotes (7,8). Cyps have been identified in every tissue studied in humans (7,8) and have been localized to the cytoplasm (1), mitochondria (9), nucleus (8,10), and endoplasmic reticulum (11) in various mammalian cells and the cytoplasm and periplasm (5,12) in E. coli. Three well characterized Cyps, CypA (18 kDa) (1,13), CypB (21 kDa) (11), and CypC (22.8 kDa) (14), are studied and discussed in this paper.…”

Section: Cyclophilins (Cyps)mentioning

confidence: 99%

Native Recombinant Cyclophilins A, B, and C Degrade DNA Independently of Peptidylprolyl cis-trans-Isomerase Activity

Montague

Hughes

Cidlowski

1997

Journal of Biological Chemistry

163

122

View full text Add to dashboard Cite

Previous work in our laboratory (Montague, J., Gaido, M., Frye, C., and Cidlowski, J. (1994) J. Biol. Chem. 269, 18877-18880) has shown that human recombinant cyclophilins A, B, and C have sequence homology with the apoptotic nuclease NUC18 and that denatured cyclophilins can degrade DNA. We have now evaluated the nucleolytic activity of recombinant cyclophilins under native conditions. We show that nuclease activity inherent to cyclophilins is distinct from cis-trans-peptidylprolyl isomerase activity and is similar to that described for apoptotic nucleases. Cyclophilin nucleolytic activity is stimulated by Ca 2؉ and/or Mg 2؉, with a combination of the two being optimal for cyclophilins A and B. Mg 2؉ alone is sufficient for cyclophilin C nuclease activity. pH optimums are in the range of pH 7.5-9.5. Cyclophilins can degrade both single-stranded and double-stranded DNA. Additionally, cyclophilins produce 3-OH termini in linear double-stranded substrates, suggesting the cuts produced are similar to those of apoptotic cells. Cyclophilins also display endonucleolytic activity, demonstrated by their ability to degrade supercoiled DNA. In the absence of ions, cyclophilins bind linearized DNA. When added to nuclei from nonapoptotic cells, cyclophilin C induces 50-kilobase pair DNA fragmentation but not internucleosomal fragmentation. Together, these data suggest that cyclophilins are involved in degradation of the genome during apoptosis.

show abstract

“…It is also possible that the inhibition ofmitochondrial PPIase is responsible for the toxic effects of cyclosporin A on liver and kidney [1,2]. In support of this hypothesis, toxicity is associated with changes in mitochondrial morphology [1,26] which are not as severe with cyclosporin G as with cyclosporin A [27,28]. FK-506 appears to lack these side-effects [29].…”

Section: Discussion Ppiase Is Involved In Pore Formationmentioning

confidence: 99%

Further evidence that cyclosporin A protects mitochondria from calcium overload by inhibiting a matrix peptidyl-prolyl cis-trans isomerase. Implications for the immunosuppressive and toxic effects of cyclosporin

Griffiths

Halestrap

1991

Biochemical Journal

212

131

View full text Add to dashboard Cite

The Ki values of cyclosporins A, G and H for the peptidyl-prolyl cis-trans isomerase (PPIase) of liver and heart mitochondria are about 2, 20 and 500 nM respectively. This parallels their profile as inhibitors of non-specific pore opening of mitochondria induced by supraphysiological Ca2+ concentrations. The novel immunosuppressant FK-506 gave little inhibition of either process at 5 microM. These data support our previous hypothesis [Halestrap & Davidson (1990) Biochem. J. 268, 153-160] that pore opening involves an interaction between matrix PPIase and the adenine nucleotide translocase. It is suggested that this model may help to clarify the mechanism of action of cyclosporin as an immunosuppressant and its toxic effects on the liver and kidney following prolonged therapy.

show abstract

Cyclosporine—relationship of Side Effects to Mode of Action

Cited by 59 publications

References 0 publications

Dynamin-2-Dependent Targeting ofMannheimia haemolyticaLeukotoxin to Mitochondrial Cyclophilin D in Bovine Lymphoblastoid Cells

Dynamin-2-Dependent Targeting ofMannheimia haemolyticaLeukotoxin to Mitochondrial Cyclophilin D in Bovine Lymphoblastoid Cells

Native Recombinant Cyclophilins A, B, and C Degrade DNA Independently of Peptidylprolyl cis-trans-Isomerase Activity

Further evidence that cyclosporin A protects mitochondria from calcium overload by inhibiting a matrix peptidyl-prolyl cis-trans isomerase. Implications for the immunosuppressive and toxic effects of cyclosporin

Contact Info

Product

Resources

About