Lithium chloride perturbation of cis-trans peptide bond equilibria: effect on conformational equilibria in cyclosporin A and on time-dependent inhibition of cyclophilin

Kofron, James L.; Kuzmič, Petr; Kishore, Vimal; Gemmecker, Gerd; Fesik, Stephen W.; Rich, Daniel H.

doi:10.1021/ja00033a047

Cited by 122 publications

(109 citation statements)

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“…This structure determination of . This is in contrast to CS, which requires 3045 min for complex formation with CYP [17,33] and supports the hypothesis that CS has to undergo structural changes before complex formation with CYP in a rate limiting step.…”

Section: Resultssupporting

confidence: 82%

The 3D structure of a cyclosporin analogue in water is nearly identical to the cyclophilin‐bound cyclosporin conformation

et al. 1994

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The conformation of [D-MeSe?-D-Ser-(O-Gly)*]CS, a water soluble cyclosporin derivative, has been determined in (D,)DMSO and in water using NMR. In these polar solvents the conformation is identical and very similar to the structure found in the cyclophilin-cyclosporin complex. However, it differs significantly from its conformation in deuterated chloroform. This demonstrates unambiguously that the large structure change is induced primarily by the polar solvent rather than by complex formation with cyclophilin.

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Section: Resultssupporting

confidence: 82%

The 3D structure of a cyclosporin analogue in water is nearly identical to the cyclophilin‐bound cyclosporin conformation

et al. 1994

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“…Walsh, unpubl.). It is likely that the mutant CyPs selectively bind the trans conformation of CsA, which exists in solution (Kessler et al, 1990;Altschuh et al, 1992) and has recently been shown to be the active species for PPIase inhibition (Kofron et al, 1992). In this view, the CyP active site serves solely as a scaffolding apparatus to present bound drug in an active conformation.…”

Section: Calcineurin Inhibition Studiesmentioning

confidence: 99%

Active site mutants of human cyclophilin A separate peptidyl‐prolyl isomerase activity from cyclosporin A binding and calcineurin inhibition

et al. 1992

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Based on recent X-ray structural information, six site-directed mutants of human cyclophilin A (hCyPA) involving residues in the putative active site--54, R55, F60, Q l l l , F113, and H126-have been constructed, overexpressed, and purified from Escherichia coli to homogeneity. The proteins W121A (Liu, J., Chen, C.-M., & Walsh, C.T., 1991a, Biochemistry30, 2306-2310), H54Q, R55A, F60A, Q l l l A , F113A, and H126Q were assayed for cis-trans peptidyl-prolyl isomerase (PPIase) activity, their ability to bind the immunosuppressive drug cyclosporin A (CsA), and protein phosphatase 2B (calcineurin) inhibition in the presence of CsA. Results indicate that H54Q, Q l l l A , F113A, and W121A retain 3-15% of the catalytic efficiency (kcoJKm) of wild-type recombinant hCyPA. The remaining three mutants (R55A, F60A, and H126Q) each retain less than 1% of the wild-type catalytic efficiency, indicating participation by these residues in PPIase catalysis. Each of the mutants bound to a CsA affinity matrix. The mutants R55A, F60A, F113A, and H126Q inhibited calcineurin in the presence of CsA, whereas W121A did not. Although CsA is a competitive inhibitor of PPIase activity, it can complex with enzymatically inactive cyclophilins and inhibit the phosphatase activity of calcineurin.

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“…The time needed to reach a stable level of CS-antibody complex is likely to reflect a conformational transition of CS. Kofron et al [7] observed different behaviours for CS initially dissolved in TFE and TFE/LiC1. In our case, readings obtained with CS dissolved in TFE/LiCI (Fig.…”

Section: Monitoring Changes In Protein-cs Interaction With Timementioning

confidence: 99%

Interaction of cyclosporin A with an Fab fragment or cyclophilin Affinity measurements and time‐dependent changes in binding

et al. 1993

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Different conformers of the immunosuppressant cyclosporin A have been observed in structural studies of the isolated molecule and of its complex with cyclophilin or with an Fab fragment. The factors that control this conformational change are not well understood. Variations in the amount of complex formed with cyclophilin or with the antibody were measured as a function of time after adding cyclosporin to the proteins, using the Pharmacia BIAcore biosensor instrument. Up to 1 hour was needed to reach maximum complex formation in solution, which is likely to reflect the time needed for a conformational transition ofcyclosporin. The equilibrium affinity constant of both proteins for cyclosporin has been measured.

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Lithium chloride perturbation of cis-trans peptide bond equilibria: effect on conformational equilibria in cyclosporin A and on time-dependent inhibition of cyclophilin

Cited by 122 publications

References 0 publications

The 3D structure of a cyclosporin analogue in water is nearly identical to the cyclophilin‐bound cyclosporin conformation

The 3D structure of a cyclosporin analogue in water is nearly identical to the cyclophilin‐bound cyclosporin conformation

Active site mutants of human cyclophilin A separate peptidyl‐prolyl isomerase activity from cyclosporin A binding and calcineurin inhibition

Interaction of cyclosporin A with an Fab fragment or cyclophilin Affinity measurements and time‐dependent changes in binding

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