HDX-MS Method Development

Conference Year

2024

Abstract

Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a powerful tool in molecular biology for studying protein structure and dynamics. This study aimed to develop an optimized HDX workflow using bovine cytochrome C (Cyto. C) as a test standard. Front end chromatography was set up to achieve in-line pepsin digestion and fast separation of the resulting peptides at 0℃. Mass spectra were analyzed using HDExaminer software to identify mass spectral shifts of peptic peptides in the deuterated Cyto. C and to calculate the corresponding percent deuterium uptake. This provided experimental evidence about the protein's structure and dynamics.

Primary Faculty Mentor Name

Ying Wai Lam

Graduate Student Mentors

Sydney Cohn Guthrie

Status

Undergraduate

Student College

College of Arts and Sciences

Program/Major

Biochemistry

Primary Research Category

Life Sciences

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HDX-MS Method Development

Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a powerful tool in molecular biology for studying protein structure and dynamics. This study aimed to develop an optimized HDX workflow using bovine cytochrome C (Cyto. C) as a test standard. Front end chromatography was set up to achieve in-line pepsin digestion and fast separation of the resulting peptides at 0℃. Mass spectra were analyzed using HDExaminer software to identify mass spectral shifts of peptic peptides in the deuterated Cyto. C and to calculate the corresponding percent deuterium uptake. This provided experimental evidence about the protein's structure and dynamics.