Rat Liver Sinusoidal Surface N-Linked Glycoproteomic Analysis by Affinity Enrichment and Mass Spectrometric Identification

---

Jianglin Li^1#, Jun Gao^2#, Miao Jiang^1#, Jia Chen¹, Zhonghua Liu¹, Ping Chen^1*, and Songping Liang^1*

¹Key Laboratory of Protein Chemistry and Developmental Biology of the Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha, 410081, P. R. China; E-mail: chenp@hunnu.edu.cn; liangsp@hunnu.edu.cn

²Qingyuan City People’s Hospital of Jinan University, Qingyuan, Guangdong, 511518, P. R. China

^# These authors contributed equally to this work.

^* To whom correspondence should be addressed.

Received April 8, 2014; Revision received November 11, 2014

---

Glycosylation in liver is one of the most biologically important protein modifications. It plays critical roles in many physiological and pathological processes by virtue of its unique location at the blood–tissue interface, including angiogenesis, liver cancer, cirrhosis, and fibrosis. To analyze glycosylation of plasma membrane proteins in liver sinusoidal endothelial cells (LSEC), N-glycopeptides of the LSEC surface were enriched using a filter-assisted sample preparation-based lectin affinity capture method and subsequently identified with mass spectrometry. In total, 225 unique N-glycosylation sites on 152 glycoproteins were identified, of which 119 (53%) sites had not previously been determined experimentally. Among the glycoproteins, 53% were classified as plasma membrane proteins and 47 (31%) as signaling proteins and receptors. Moreover, 23 cluster of differentiation antigens with 49 glycopeptides were detected within the membrane glycoproteins of the liver sinusoidal surface. Furthermore, bioinformatics analysis revealed that the majority of identified glycoproteins have an impact on processes of LSEC. Therefore, N-glycoproteomic analysis of the liver sinusoidal surface may provide useful information on liver regeneration and facilitate liver disease diagnosis.

---

KEY WORDS: liver sinusoidal endothelial surface, N-glycoproteomic, N-glyco-FASP, mass spectrometry

DOI: 10.1134/S0006297915030025

---