[Sample pre-processing for mass spectrometric analysis of glycoprotein structure].

Abstract

A novel, simple and fast method based on a quadrupole ion trap (QIT) and a time-of-flight mass spectrograph was developed for the glycosylation site and glycan structure analysis of glycoprotein by in-solution digestion and in-gel digestion which did not need to enrich and label the glycopeptide. Horseradish peroxidase (HRP) and RNase B were utilised to optimize the conditions of sample pre-processing. The effects of the conditions were investigated separately and the optimal conditions were: heat denaturation of RNase B, chemical denaturation of HRP, RNase B digestion with endoproteinase Lys-C, HRP digestion with trypsin, 12 - 16 h of digestion, 50% acetonitrile-5% trifluoroacetic acid of the extraction solution, sandwich spotting method of the sample.

• Authors: Wei Zhou, Xiaohui Liu, Xinwen Zhou, Huali Shen, Pengyuan Yang
• Journal: Se pu = Chinese journal of chromatography (Se Pu) Vol. 25 Issue 5 Pg. 623-7 (Sep 2007) ISSN: 1000-8713 [Print] China
• PMID: 18161309 (Publication Type: English Abstract, Journal Article)

Chemical References

• Glycoproteins
• Horseradish Peroxidase
• Ribonucleases
• ribonuclease B
• Trypsin
• Metalloendopeptidases
• peptidyl-Lys metalloendopeptidase

Topics

• Glycoproteins (chemistry, metabolism)
• Horseradish Peroxidase (metabolism)
• Mass Spectrometry (methods)
• Metalloendopeptidases (metabolism)
• Ribonucleases (metabolism)
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (methods)
• Trypsin (metabolism)