surface enhanced raman optical activity as an ultra sensitive tool for ligand binding analysis表面增强拉曼光活动作为配体的超敏感的工具结合分析.pdfVIP

Spectroscopy 21 (2007) 143–149 143 IOS Press Surface enhanced Raman optical activity as an ultra sensitive tool for ligand binding analysis Christian Johannessen and Salim Abdali ∗ Quantum Protein Centre, QuP, Department of Physics, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark Abstract. The Surface Enhanced Resonance Raman Scattering (SERRS) and Surface Enhanced Resonance Raman Optical Activity (SERROA) spectra of myoglobin and the myoglobin–azide complex were measured on very dilute samples (100 nM protein) in order to analyze the sensitivity of SERROA spectroscopy when inducing small structural changes. While the SERRS spectra of the two compounds were virtually identical, comparison of the SERROA spectra revealed several differences, includ- ing frequency shifts and changes in signal intensity, consistent with structural change in the porphyrin prosthetic group of the protein upon azide complexation. Application of this method allows for rapid analysis of ligand binding in metalloproteins in dilute aqueous solution and could in the future, when combined with theoretical studies, increase the obtainable structural resolution of proteins beyond that of X-ray analysis. Keywords: Spectroscopic analysis, colloid, chiral, SERRS, SEROA, metalloprotein 1. Introduction A major concern in analysis of biological samples in spectroscopic chemical research, as well as in clinical or industrial applications, is sample concentration. While biological assays often are optimized to measure very dilute samples [1], or in complex mixtures such as a living cell [2], spectroscopic analysis frequently requires pure samples of concentrations close to saturation [3]. Additionally, in order to obtain a high level of detail, e.g. studying structure and function through spectroscopic techniques, long acquisition times are usually required.