JoVE Journal

Biochemistry

This content is Open Access.

Glycoproteomics of the Extracellular Matrix: A Method for Intact Glycopeptide Analysis Using Mass Spectrometry

12.2K Views

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14:02 min

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April 21st, 2017

DOI :

10.3791/55674-v

April 21st, 2017

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Javier Barallobre-Barreiro¹, Ferheen Baig¹, Marika Fava¹, Xiaoke Yin¹, Manuel Mayr¹

¹King's British Heart Foundation Centre, King's College London

Summary

This paper describes a methodology to prepare cardiovascular tissue samples for MS analysis that allows for (1) the analysis of ECM protein composition, (2) the identification of glycosylation sites, and (3) the compositional characterization of glycan forms. This methodology can be applied, with minor modifications, to the study of the ECM in other tissues.

Explore More Videos

Glycoproteomics

Extracellular Matrix

Mass Spectrometry

Tissue Fibrosis

Cardiovascular Disease

Glycopeptide Analysis

Glycosylation Site Occupancy

Chapters in this video

0:05

Title

1:05

Extraction of Extracellular Matrix Proteins

3:52

Protein Precipitation

4:57

Sequential Deglycosylation for the Assessment of N-glycosylation Site Occupancy

6:05

Peptide Cleanup Using C18 Column

8:32

Glycopeptide Enrichment

11:32

Results: Glycopeptide Extraction and Analysis

13:18

Conclusion

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