Abstract
Metallothionein has been implicated in a number of functions, including toxic metal detoxification, as a metal chaperone and in metal ion homeostasis. In this paper, we demonstrate that the beta domain of human metallothionein 1a, well-known to bind three Zn(2+) or Cd(2+) ions with nine cysteinyl sulfurs, is also capable of binding an additional Cd(2+) ion, leading to the formation of the supermetalated Cd(4)-beta-rhMT 1a. This intermediate, either by itself or in concert with the alpha domain of human metallothionein, is a likely model for metal exchange with the apoenzyme, which is one of the key roles of metallothionein. Through electrospray ionization (ESI) mass spectrometry and circular dichroism (CD) and ultraviolet (UV) spectroscopy, we show that the addition of 4.4 molar equiv of CdSO(4) to a solution of Cd(3)-beta-rhMT 1a leads to the complete conversion to Cd(4)-beta-rhMT 1a. ESI mass spectrometry was used to determine the exact speciation of beta-rhMT 1a. While the UV absorption spectrum increased slightly, the CD spectrum of Cd(4)-beta-rhMT 1a showed significant changes with the appearance of a sharp monophasic peak at 252 nm in contrast to the derivative-shaped envelope of the Cd(3)-beta-rhMT 1a species [peak extrema at (+)262 and (-)236 nm], indicating disruption of the exciton coupling in the metal-thiolate cluster. Additionally, both direct and indirect (113)Cd nuclear magnetic resonance (NMR) spectra of the Cd(3)-beta-rhMT 1a and Cd(4)-beta-rhMT 1a species were recorded. The (113)Cd NMR spectrum of Cd(4)-beta-rhMT 1a contained four cadmium peaks in the tetrahedral thiolate region at 688.8, 650.3, 635.9, and 602.5 ppm. This represents the first report of both NMR data for isolated Cd(3)-beta-rhMT 1a and supermetalated Cd(4)-beta-rhMT 1a.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.