Glycoproteins analysis

E. F. Hounsell (Ed.), Methods in Molecular Biology, Vol. 14, Glycoprotein Analysis in Biomedicine. Humana Press, 1993. 

A. Dell, J. E. Thomas-Oates, M. E. Rogers, and P. R. Tiller, Novel fast atom bombardment procedures for glycoprotein analysis, Biochimie, 70 (1988) 1435-1444. 

Orr GA, Han EK, Browne PC, et al. Identification of the major phosphorylation domain of murine mdrlb P-glycoprotein. Analysis of the protein kinase A and protein kinase C phosphorylation sites. J Biol Chem 1993 268(33) 25054-25062. 

The use of ConA affinity chromatography in SDS has proved to be very useful for glycoprotein analysis because it combines complete dissociation of the glycoproteins from associated components with a rapid, specific, and high yield method of isolation. Most importantly, the glycoproteins reform the complex after displacement of SDS by nonionic detergent (Bando et al., 1996 Li et al., 1999). 

Gates, R., Rathbone, E., Masterson, L., Wright, I., and Electricwala, A. 2004. Glycoprotein Analysis Manual, 1st ed., Sigma-Aldrich Co. St Louis, MO. 

Fig. 5. The glycoprotein analysis workflow proposed by Renkonen et al,43 As a proof of the principle, the authors showed an example of comprehensive approach for glycoproteomics of purified human plasma serotransferrin.

While the discussion in this text focusses on the use of LC-MS, on-line capillary electrophoresis-MS is another important tool in glycopeptide and glycoprotein analysis, as shown by the data reviewed by Zamfir and Peter-Katahnic [89]. 

Tao, W.-J. M. A. Quan, Gritsenko, D. G. Camp, M. E. Monreo, R. J. Moore, and R. D. Smith. 2005. Human plasmaN-Glycoprotein analysis by immunoaffinity substraction, hydrazine chemistry, and mass spectrometry. J. Proteom. Res. 4, 2070-2080. 

C-Linked glycosylation was first identified on ribonuclease 2 and is characterized by a-mannose (Man) attached to the 2-position of the indole side chain of tryptophan residues (Figure 6(b)). This modification has subsequently been identified on several proteins such as MUC5AC and MUC5B/ thrombospondin, and even the Ebola virus soluble glycoprotein. Analysis of protein databases for the putative C-linked mannosyla-tion consensus sequence, TrpXaaXaaTrp, suggests that a variety of proteins may bear this modification. The main role of C-linked mannosylation on protein remains to be determined. 

Methods for Glycoprotein Analysis Using Liquid Chromatography and Capillary Electrophoresis/Electrospray Mass Spectrometry... 

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