Document Type

Article

Publication Date

9-1-2015

Publication Title

Glycobiology

Abstract

Sialic acids (SAs) are widely expressed on immune cells and their levels and linkages named as sialylation status vary upon cellular environment changes related to both physiological and pathological processes. In this study, we performed a global profiling of the sialylation status of macrophages and their release of SAs in the cell culture medium by using flow cytometry, confocal microscopy and liquid chromatography tandem mass spectrometry (LC-MS/MS). Both flow cytometry and confocal microscopy results showed that cell surface α-2,3-linked SAs were predominant in the normal culture condition and changed slightly upon treatment with atorvastatin for 24 h, whereas α-2,6-linked SAs were negligible in the normal culture condition but significantly increased after treatment. Meanwhile, the amount of total cellular SAs increased about three times (from 369 ± 29 to 1080 ± 50 ng/mL) upon treatment as determined by the LC-MS/MS method. On the other hand, there was no significant change for secreted free SAs and conjugated SAs in the medium. These results indicated that the cell surface α-2,6 sialylation status of macrophages changes distinctly upon atorvastatin stimulation, which may reflect on the biological functions of the cells.

Comments

This work was supported by Faculty Research Development Fund and the research fund from the Center for Gene Regulation in Health and Disease (GRHD) at Cleveland State University supported by Ohio Department of Development (ODOD). The authors acknowledge the National Science Foundation Major Research Instrumentation Grant (CHE-0923398) for supporting Q-Trap 5500 mass spectrometer instrument, the National Institution of Health for supporting Nikon A1Rsi confocal microscope (1S10OD010381). This work was partially supported by grants from The National Natural Science Foundation of China (31328006). H Nie appreciates the China Oversea Scholar Award from China Scholarship Council.

DOI

10.1093/glycob/cwv038

Version

Postprint

Volume

25

Issue

9

Included in

Chemistry Commons

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