Native electrospray and electron-capture dissociation in FTICR mass spectrometry provide top-down sequencing of a protein component in an intact protein assembly

  • Hao Zhang
  • Weidong Cui
  • Jianzhong Wen
  • Robert E. Blankenship
  • Michael L. Gross
Short Comunication

Abstract

The intact yeast alcohol dehydrogenase (ADH) tetramer of 147 kDa was introduced into a FTICR mass spectrometer by native electrospray. Electron capture dissociation of the entire 23+ to 27+ charge state distribution produced the expected charge-reduced ions and, more unexpectedly, 39 c-type peptide fragments that identified N-terminus acetylation and the first 55 amino acids. The results are in accord with the crystal structure of yeast ADH, which shows that the C-terminus is buried at the assembly interface, whereas the N-terminus is exposed, allowing ECD to occur. This remarkable observation shows promise that a top-down approach for intact protein assemblies will be effective for characterizing their components, inferring their interfaces, and obtaining both proteomics and structural biology information in one experiment.

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Copyright information

© American Society for Mass Spectrometry. Published by Elsevier Inc 2010

Authors and Affiliations

  • Hao Zhang
    • 1
  • Weidong Cui
    • 1
  • Jianzhong Wen
    • 1
  • Robert E. Blankenship
    • 1
  • Michael L. Gross
    • 1
  1. 1.Department of ChemistryWashington University in St. LouisSt. LouisUSA

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