XAFScourse6a 同步辐射应用新进展课程.pptVIP

SR Research Strengths Structural Biology X-ray Crystallography Solution and solid-state NMR Spectroscopy Single-molecule Ultra-fast X-ray Absorption Computational Methods Protein folding, dynamics, structure prediction Regulatory Networks Spectroscopically quiet metals The Role of Zn in Biology Zinc in Biology Zinc is the most common “trace” element Zinc is the only metal that is known to be required by at least one enzyme in every major class of reactivity Zinc levels are tightly regulated Total Zinc (from E. coli to human cells) is ca. 10–4 M Free Zinc is estimated at 1 ion / cell Why should biology want to use Zn? Good bio-availability Redox inert (safe in presence of O2) Thermodynamically stable, e.g., Zn(SR)42– Rapid ligand exchange Good Lewis acid d10 configuration No geometric preference Coordination sphere can easily be expanded Biological Zn sites Homocysteine MetE (cobalamin independent MetSyn) contains Zn XANES spectra are sensitive to ligation Zn EXAFS is remarkably insensitive to changes in ligation. Zn-S and Zn-N EXAFS signals are approximately out of phase Threshold energy changes apparent ligation The Zn site in MetE (cobalamin independent MetSyn) has ZnS2(O/N)2 ligation. XANES spectra change on addition of homocysteine. Changes in ligation are due to homocysteine binding to Zn Se EXAFS confirms structural picture of MetE and MetH sites Combination of Zn + Se EXAFS consistent with small distortions from tetrahedral geometry in substrate-bound enzyme Mechanism of homocysteine methylation involves both Co and Zn Epoxide carboxylase Substrate binding affects the Zn site in epoxide carboxylase . . . …consistent with SCoM binding to Zn Note that native and native+CoM+epoxide have distinct edges despite having identical EXAFS Possible roles for Zn in epoxide carboxylase Protein Farnesyl transferase Protein Farnesyl transferase Zn is 4-coordinate;peptide sulfur binds only in ternary complex Se EXAFS confirms displacement of product from Zn Carboxyla