Green algae Chlamydomonas reinhardtii possess endogenous sialylated N-glycans

FEBS Open Bio. 2011 Oct 30:1:15-22. doi: 10.1016/j.fob.2011.10.003. Print 2011 Dec.

Abstract

Green algae have a great potential as biofactories for the production of proteins. Chlamydomonas reinhardtii, a representative of eukaryotic microalgae, has been extensively used as a model organism to study light-induced gene expression, chloroplast biogenesis, photosynthesis, light perception, cell-cell recognition, and cell cycle control. However, little is known about the glycosylation machinery and N-linked glycan structures of green algae. In this study, we performed mass spectrometry analysis of N-linked oligosaccharides released from total extracts of Chlamydomonas reinhardtii and demonstrated that C. reinhardtii algae possess glycoproteins with mammalian-like sialylated N-linked oligosaccharides. These findings suggest that C. reinhardtii may be an attractive system for expression of target proteins.

Keywords: 2-AB, 2-aminobenzamide; CST, CMP-sialic acid transporter; Chlamydomonas reinhardtii; ER, endoplasmic reticulum; HPLC-FLD, high-performance liquid chromatography using fluorescence detection; HRP, horseradish peroxidase; MALDI-TOF-MS, matrix-assisted laser desorption/ionization time of flight mass spectrometry; MS, mass spectrometry; Mass spectrometry analysis; N-Glycan structure; PBS, phosphate buffered saline; PVDF, polyvinylidene fluoride; RCA, Ricinus communis agglutinin; SDS–PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; Sialic acid; TBS, Tris-buffered saline; hEPO, human erythropoietin.