Pb2+ as modulator of protein-membrane interactions

Krystal A. Morales, Mauricio Lasagna, Alexey V. Gribenko, Youngdae Yoon, Gregory D. Reinhart, James C. Lee, Wonhwa Cho, Pingwei Li, Tatyana I. Igumenova

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

Lead is a potent environmental toxin that mimics the effects of divalent metal ions, such as zinc and calcium, in the context of specific molecular targets and signaling processes. The molecular mechanism of lead toxicity remains poorly understood. The objective of this work was to characterize the effect of Pb2+ on the structure and membrane-binding properties of C2α. C2α is a peripheral membrane-binding domain of Protein Kinase Cα (PKCα), which is a well-documented molecular target of lead. Using NMR and isothermal titration calorimetry (ITC) techniques, we established that C2α binds Pb2+ with higher affinity than its natural cofactor, Ca2+. To gain insight into the coordination geometry of protein-bound Pb2+, we determined the crystal structures of apo and Pb2+-bound C2α at 1.9 and 1.5 Å resolution, respectively. A comparison of these structures revealed that the metal-binding site is not preorganized and that rotation of the oxygen-donating side chains is required for the metal coordination to occur. Remarkably, we found that holodirected and hemidirected coordination geometries for the two Pb2+ ions coexist within a single protein molecule. Using protein-to-membrane Förster resonance energy transfer (FRET) spectroscopy, we demonstrated that Pb 2+ displaces Ca2+ from C2α in the presence of lipid membranes through the high-affinity interaction with the membrane-unbound C2α. In addition, Pb2+ associates with phosphatidylserine- containing membranes and thereby competes with C2α for the membrane-binding sites. This process can contribute to the inhibitory effect of Pb2+ on the PKCα activity.

Original languageEnglish (US)
Pages (from-to)10599-10611
Number of pages13
JournalJournal of the American Chemical Society
Volume133
Issue number27
DOIs
StatePublished - Jul 13 2011

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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    Morales, K. A., Lasagna, M., Gribenko, A. V., Yoon, Y., Reinhart, G. D., Lee, J. C., Cho, W., Li, P., & Igumenova, T. I. (2011). Pb2+ as modulator of protein-membrane interactions. Journal of the American Chemical Society, 133(27), 10599-10611. https://doi.org/10.1021/ja2032772