Metallothionein

1. cordblood exon 2 exon 1 exon 3 5’ 3’ UTR intron 1 intron 2 UTR Ex vivo expanded RBC precursors: without Zn Ex vivo expanded RBC precursors: 100 µM of Zn Biochemistry, Molecular and Structural Biology • Low Mr (6000 – 7000) • High metal content (7 M2+ or 12 M1+) • 30% Cys (20/61 amino acids, Cys-X-Cys) • Metal-thiolate clusters • Human chromosome 16q13 • 10 functional and 7 non functional MT isoforms • Structure – function relationship • Interference with the signal transduction path: • Mutants / peptides • Antibodies (mono- and polyclonal) • Anti-idiotypic antibodies (monoclonal) • Receptor fragments • Anti-sense technology • Over-expression – transfection • Natural inhibitors • homeostasis of essential heavy metals (Zn) • detoxification of toxic heavy metals (Cd) • drug resistance to alkylating agents • endogenous anti-mutagenic agents • defense against oxidative stress • possible role in differentiation Metallothionein Interferon-g Molecular Biology&Signal Transduction Techniques Current research • Basic biochemical, physical chemical and molecular biological techniques • Protein & DNA sequencing • Atomic absorption (AAS) • Fluorescence microscopy • Radiaoctive labeling • HPLC & FPLC • ELISA • PCR • Prokaryotic and eukaryotic cell cultures • Bioluminescence • Cell fractionation (e.g. MACS) • Cytospin • Immunocytochemistry • Transformation and transfection • Electroporation and electrofusion • Biosynthesis of MT in precursor cells from cord blood and in cell lines • Characterisation of MT from a-nucleated blood cells • Interaction of MT with nucleic acids: nucleolytic effect • Over-expression of MT in eukaryotic cells (cell lines) http://www.chem.kuleuven.ac.be/research/bio/mbsten.html Laboratory of Biochemistry Prof. Marc DE LEY Group Molecular Biology and Signal TransductionCelestijnenlaan 200G (2nd floor) Leuven (Heverlee)