1. PI: Kuo Kan Liang 梁國淦 Annual Report DiCK / SLaBS RCAS, Academia Sinica RCAS, AS and BST, NTU

2. Theme of Researches Chemical reactions and transport processes in biological systems – Special emphases: Membrane protein ion channels Methodology: – Spectroscopic methods – Electrochemistry methods – Both include theory and experiments RCAS, AS and BST, NTU

3. Methodology: Spectroscopic Methods Eternal goal: Spectroscopic observation of the kinetics with as little labeling as possible IR spectroscopy; – Mid-IR range observation of amide-I peaks – Ultrafast multiphoton coherent spectroscopy such as 2D-IR – Time-resolved FT-IR Fluorescence spectroscopy: – Natural amino acids: Tryptophan, tyrosine RCAS, AS and BST, NTU

4. Methodology: Spectroscopic Methods Protein backbone modes couplings can be observed as vibration coherence as well as mode anharmonicity Past achievements – Theory of anharmonic effect on absorption lineshape – Theory of effect of near-by vibronic level on the 3- pulse photon echo signal of a vibration mode – Theory of anharmonic and mode-mode coupling effect on the 3-pulse photon echo signal of a vibration mode. RCAS, AS and BST, NTU

5. Publications: Spectroscopic Methods Chu, Z.*; Liang, K. K.*; Hayashi, M.; Lin, S. H., “Theoretical treatment of anharmonic effect on molecular absorption, fluorescence spectra, and electron transfer”, Chem. Phys. 2009, 358, 137. RCAS, AS and BST, NTU

6. Publications: Spectroscopic Methods Villaeys, A. A.*; Dappe, Y.; Liang, K. K.; Lin, S. H., “Influence of neighboring levels in three-pulse photon-echo processes”, Phys. Rev. A 2009, 79, 053418. (Virtual Journal of Ultrafast Science 2009, Vol. 8, Issue 6) RCAS, AS and BST, NTU

7. Publications: Spectroscopic Methods Liang, K. K.*; Villaeys, A. A., “Anharmonicity and mode-mode coupling effects analyzed by 4WM photon-echo spectroscopy”, Phys. Rev. A 2010 (Accepted with minor corrections). RCAS, AS and BST, NTU

8. Methodology: Electrochemistry Methods Impedance spectroscopy (transfer function) of ion channels Modified electrophysiology (patch-clamp) experiments – Patch-clamp impedance spectroscopy – Patch-Clamp fluorometry (Electrophysiology + Spectroscopy) – Time-resolved experiments: synchronization method

9. Publication: Electrophysiology Methods Chang, H.-K.; Martin, L. J.; Liang, K. K.; Shieh, R.-C.*, “K+ binding in G-loop and water cavity facilitates Ba 2+ movement in the Kir2.1 Channel”, Biochim. Biophys. Acta 2009, 1788, 500. RCAS, AS and BST, NTU Hill plot: Hill coefficient ~ 2.5 Indicating that cooperative binding is not the sole story

10. Further Developments: Short term Theory of kinetics of cooperative systems * Stochastic model of ion transport in channels * Molecular dynamics simulations and analysis of hydrated cations transport in bulk and in artificial channels Patch-clamp fluorometry of unlabeled Kir 2.1/ 2.2 channels RCAS, AS and BST, NTU

11. Further Developments: Mid-term Impedance of ion channels – Theoretical method: Chemical kinetics simulation of the ion transport process in ion channels – Experimental method: AC patch-clamp Advanced materials for CO 2 separation – Theory of CO 2 or CO 3 2– transport in pores such as in polymer-metal framework (PMF) – Model system construction for cell-free CO 2 reduction using enzymes RCAS, AS and BST, NTU

12. Further Developments: Long-term Thermodynamics and kinetics of single- / few- biological-macromolecule events – Theory of reactive transport in pores / channels – Thermodynamics and kinetics of binding in channels – Thermodynamics systems with electrostatic works Electrochemical and structural kinetics of cells – Synchronous observations of reactions, transport processes, and structural changes on membrane – Engineering of cellular or cell-free molecular factories RCAS, AS and BST, NTU