Detection of conformation types of cyclosporin retaining intramolecular hydrogen bonds by mass spectrometry.

Suk-Joon Hyung, Xidong Feng, Ye Che, Justin G Stroh, Michael Shapiro

Index: Anal. Bioanal. Chem 406(24) , 5785-94, (2014)

Full Text: HTML

Abstract

Cyclosporin is a family of neutral cyclic undecapeptides widely used for the prevention of organ transplant rejection and controlling viral infection. The equilibrium of conformations assumed by cyclosporin A in response to the solvent environment is thought to play a critical role in enabling good membrane penetration, which improves upon shielding the polarity of the molecule through forming intramolecular hydrogen bonds. However, the distribution of structures and their internal hydrogen bond geometries have not been elucidated thus far across the series of cyclosporins. Herein, we elucidate the conformational heterogeneity of cyclosporins using a set of analytical approaches including ion mobility mass spectrometry, hydrogen-deuterium exchange, and molecular dynamics simulation. Ion mobility measurements reveal a specific conformational distribution for each cyclosporin derivative in a structure-dependent manner. In general, we observe that the more compact conformer is associated with a greater frequency of intramolecular hydrogen bonds. Cyclosporin A is populated by structures with an extensive hydrogen bond network that is lacking in cyclosporin H, which is composed predominantly of a single compact conformation. The slower dynamics of cyclosporin H backbone is also consistent with the lack of hydrogen bonds. Furthermore, we find a strong correlation between the steric bulk of the side chain at position 2 of cyclosporin and the distribution of conformers due to differential accommodation of side chains within the macrocycle, and also report a wide range of conformational dynamics in solution.