Direct quantification of protein glycan phosphorylation

Phosphorylation is an important post-translational modification (PTM) of proteins and a critical quality attribute for protein therapeutics, especially if it is required for protein function or sub-cellular targeting. Most current methods to quantify phosphorylation are time-consuming, indirect, or...

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Veröffentlicht in:BioTechniques. - 1993. - 63(2017), 3 vom: 01. Sept., Seite 117-123
1. Verfasser: Ketcham, Stephanie A (VerfasserIn)
Weitere Verfasser: Ashraf, Muhammad, Madhavarao, Chikkathur N
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2017
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. glycosylation lysosomal enzymes mannose-6-phosphate phospho-glycans phosphorylation 1,3-bis(bis(pyridin-2-ylmethyl)amino)propan-2-ol Amino Acids mehr... Polysaccharides Pyridines Glucuronidase EC 3.2.1.31 Cathepsin D EC 3.4.23.5
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520 |a Phosphorylation is an important post-translational modification (PTM) of proteins and a critical quality attribute for protein therapeutics, especially if it is required for protein function or sub-cellular targeting. Most current methods to quantify phosphorylation are time-consuming, indirect, or require specific instrumentation and technical skills. Here, we report the adaptation of a phosphate-specific binding dye and common laboratory instruments for quantification of relative amounts of phosphorylated glycans as well as phosphorylation of amino acid residues on the backbones of proteins. Our results show that quantification of phosphorylation using the new method agrees with published data on the number of phosphorylated glycosylation sites for two lysosomal enzymes: β-glucuronidase (GUS) and cathepsin D 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a Research Support, U.S. Gov't, P.H.S. 
650 4 |a glycosylation 
650 4 |a lysosomal enzymes 
650 4 |a mannose-6-phosphate 
650 4 |a phospho-glycans 
650 4 |a phosphorylation 
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650 7 |a Cathepsin D  |2 NLM 
650 7 |a EC 3.4.23.5  |2 NLM 
700 1 |a Ashraf, Muhammad  |e verfasserin  |4 aut 
700 1 |a Madhavarao, Chikkathur N  |e verfasserin  |4 aut 
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