Measurement of Submillisecond Protein Folding Using Trp Fluorescence and Photochemical Oxidation

David Witalka; Lisa J Lapidus

doi:10.1007/978-1-0716-1716-8_7

Measurement of Submillisecond Protein Folding Using Trp Fluorescence and Photochemical Oxidation

Methods Mol Biol. 2022:2376:135-142. doi: 10.1007/978-1-0716-1716-8_7.

Authors

David Witalka¹, Lisa J Lapidus²

Affiliations

¹ Department of Physics and Astronomy, Michigan State University, East Lansing, MI, USA.
² Department of Physics and Astronomy, Michigan State University, East Lansing, MI, USA. lapidus@msu.edu.

PMID: 34845607
DOI: 10.1007/978-1-0716-1716-8_7

Abstract

Observation of protein folding on submillisecond time scales requires specialized ultra-rapid mixers coupled to optical or chemical probes. Here we describe the protocol for employing a microfabricated mixer with a mixing time of 8 μs coupled to a UV confocal microscope. This instrument can detect Trp fluorescence and also excite hydroxyl radicals that label the folding protein which can be detected by mass spectrometry.

Keywords: Fast photochemical oxidation of proteins (FPOP); Microfluidic mixer; Protein folding; Trp fluorescence.

MeSH terms

Hydroxyl Radical
Mass Spectrometry
Oxidation-Reduction
Protein Folding*

Substances

Hydroxyl Radical