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Sequencing Proteins from Bottom to Top: Combining Techniques for Full Sequence Analysis of Glucokinase

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Abstract

Proteomics-based mass spectrometry has gained increasing amounts of popularity in recent years. In particular, high resolution accurate mass measurements in mass spectrometry has gained notoriety for giving the capability of high throughput analysis with lower cost to the user. In particular, its uses in the identification of protein sequence through the utilization of bottom-up, middle-down, and top-down approaches has been widely discussed. In this chapter, we discuss the advantages of each technique as well as using the techniques in tandem to gain well-rounded structural data on our protein of interest, glucokinase. The study will focus on the use of Fourier-transform ion cyclotron resonance (FT-ICR) mass spectrometry, but give insights into the advantages that may come from the utilization of other high resolution techniques.

Original languageEnglish
Pages (from-to)111-119
Number of pages9
JournalAdvances in experimental medicine and biology
Volume1140
DOIs
StatePublished - 2019

Keywords

  • Bottom-up
  • Electrospray ionization
  • Enzymatic digestion
  • Proteomics
  • Sequencing
  • Tandem mass spectrometry
  • Top-down

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