Glycosylation and secretion of human α-amylases

Shou Takashima, Junko Amano

Laboratory of Glycobiology, The Noguchi Institute, Tokyo, Japan.
New Industry Creation Hatchery Center, Tohoku University, Sendai, Japan.

DOI: 10.4236/abc.2012.21002                

Abstract

Three human α-amylases exist: Amy1 (salivary amylase), Amy2A (pancreatic amylase), and Amy2B (expressed in various tissues). These amylases share a 97% - 99% amino acid sequence identity, and two potential N-glycosylation sites (N427 and N476) are commonly found in the C-terminal region. In general, salivary amylase is more frequently glycosylated than pancreatic amylase, and it is still uncertain why differences in the glycosylation pattern among human amylase iso-zymes occur. In this study, we found that there was no significant change of ratio of glycosylated molecules among isozymes produced by the same cultured cells, indicating that glycosylation of amylase is influenced by the type of cell producing the enzyme rather than being an inherent property of the amylase isozymes. We analyzed the glycosylation efficiency of N-glycosylation sites in recombinant Amy2A mutants produced by HEK293 cells and found that glycosylation efficiencies of N427 and N476 were 3% - 18% and 40% - 52%, respectively, indicating that the major N-glycosylation site of glycosylated Amy2A produced by HEK293 cells is N476. The difference in the glycosylation efficiency of each N-glycosylation site also seemed to contribute in part to generate different glycosylation patterns of human amylases. We also confirmed that the C-terminal region of human amylase plays a critical role in secretion, although glycosylation does not play a part in this effect.

Keywords

Amylase; Gycosylation Efficiency; N-Glycan; Secretion

Conflicts of Interest

The authors declare no conflicts of interest.

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