Improved stopped‐flow time‐resolved resonance Raman spectroscopy device for studying enzymatic reactions. (10th February 2017)

• Record Type: Journal Article
• Authors: Yanagisawa, Sachiko; Deshpande, Megha Subhash; Hirota, Shun; Nakagawa, Tatsuo; Ogura, Takashi
• Is Part Of: Journal of Raman spectroscopy. Volume 48:Number 5(2017)
• Page Start: 680
• Page End: 685
• Publication Date: 2017-02-10
• Subjects: Raman spectroscopy -- Periodicals; 535.846
• DOI: 10.1002/jrs.5100
• Languages: English
• ISSNs: 0377-0486

Abstract

Abstract : An improved stopped‐flow resonance Raman spectroscopy device was constructed using a stopped‐flow mixer with a dead time of 3 ms and a mixing volume of 0.1 mL. The device was tested using myoglobin, where the formation reaction of a high‐valent heme species, ferryl‐oxo heme, was monitored by time‐resolved resonance Raman spectroscopy after mixing a ferric myoglobin solution with a hydrogen peroxide solution. The ferryl‐oxo heme formation rate constant obtained by Raman spectroscopy is in good agreement with the rate constant obtained by conventional stopped‐flow absorption spectroscopy for the same reaction under the same conditions. It is proved by these results that the present device is generally applicable to enzyme–substrate reactions with a significantly higher time resolution than previously reported. Copyright © 2017 John Wiley & Sons, Ltd. Abstract : An improved stopped‐flow resonance Raman spectroscopy device was constructed using a stopped‐flow mixer with a dead time of 3 ms and a mixing volume of 0.1 mL. The device was tested using myoglobin, where the formation reaction of a high‐valent heme species, ferryl‐oxo heme, was monitored by time‐resolved resonance Raman spectroscopy after mixing a ferric myoglobin solution with a hydrogen peroxide solution. The present device is generally applicable to enzyme–substrate reactions with a significantly higher time resolution than previously reported.