title: Pb-207 NMR spectroscopy reveals that Pb(II) coordinates with glutathione (GSH) and tris cysteine zinc finger proteins in a PbS3 coordination environment.
abstract: 207Pb NMR spectroscopy can be used to monitor the binding of Pb(II) to thiol rich biological small molecules such as glutathione and to zinc finger proteins. The UV/visible (UV/Vis) absorption band centered at 334 nM and the observed 207Pb-signal in 207Pb NMR (δ~5750 ppm) indicate that glutathione binds Pb(II) in a trigonal pyramidal geometry (PbS3) at pH 7.5 or higher with a 1:3 molar ratio of Pb(II) to GSH. While previous studies using UV/Vis and extended X-ray absorption fine structure (EXAFS) spectroscopy were interpreted to show that the zinc binding domain from HIV nucleocapsid protein (HIV-CCHC) binds Pb(II) in a single PbS3 environment, the more sensitive 207Pb NMR spectra (at pH 7.0, 1:1 molar ratio) provide compelling evidence for the presence of two PbS3 structures (δ=5790 and 5744 ppm), one of which is more stable at high temperatures. It has previously been proposed that the HIV-CCHH peptide does not fold properly to afford a PbS2N motif, because histidine does not bind to Pb(II). These predictions are confirmed by the present studies. These results demonstrate the applicability of 207Pb NMR to biomolecular structure determination in proteins with cysteine binding sites for the first time.
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.
authors.forename: Kosh P
authors.initials: KP
authors.lastname: Neupane
authors.forename: Vincent L
authors.initials: VL
authors.lastname: Pecoraro
mesh_terms: D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D005978:Glutathione / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007854:Lead / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D019590:Nucleocapsid Proteins / Q000737:chemistry*; D013696:Temperature; D016335:Zinc Fingers
chemical_list: D019590:Nucleocapsid Proteins; D007854:Lead; D005978:Glutathione; D003545:Cysteine