Methods for the kinetic analysis of peptide N-glycosidase
The peptide N-glycanase(PNGase,also known as NGLY1 in human)is an ubiquitous enzyme found in nearly all living cells.PNGase,which is capable of catalyzing the cleavage of the glycosidic bond between asparagine and N-glycans in glycoproteins or glycopeptides,has been widely applied as valuable tool enzyme in glycoproteomics research.In addition,NGLY1 plays a crucial role in the process of endoplasmic reticulum-associated degradation,and deficiency of NGLY1 may lead to congenital disorder of glycosylation.The investigation of enzymatic reaction kinetics is of great value for the discovery or directed evolution of more abundant tool enzymes suitable for various analysis strategies,examining the correlation between NGLY1 mutants and diseases,as well as exploring the mechanisms of related disease and developing strategies for their treatment.In this paper,the discovery,physiological and pathological functions and the application of PNGase in glycoproteomics are described.Kinetic analysis methods of PNGase,including colorimetric method,high performance liquid chromatography methods,sodium dodecyl sulfate-capillary gel electrophor-esis methods,and liquid chromatography-mass spectrometry methods,are emphasized,and the advantages and disadvantages of the above methods are summarized.It provides inspiration for future advancements in the development of simpler and more sensitive kinetic analysis method for PNGase.
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维普
