Mapping N-linked Glycosylation Sites in the Secretome and Whole Cells of Aspergillus niger Using Hydrazide Chemistry and Mass Spectrometry

Journal Article · Sun Jan 01 00:00:00 EST 2012 · Journal of Proteome Research

DOI:https://doi.org/10.1021/pr200916k· OSTI ID:1035423

Wang, Lu; Aryal, Uma K; Dai, Ziyu; Mason, Alisa C; Monroe, Matthew E; Tian, Zhixin; Zhou, Jianying; Su, Dian; Weitz, Karl K; Liu, Tao; Camp, David G; Smith, Richard D; Baker, Scott E; Qian, Weijun

Protein glycosylation is known to play an essential role in both cellular functions and the secretory pathways; however, little information is available on the dynamics of glycosylated N-linked glycosites of fungi. Herein we present the first extensive mapping of glycosylated N-linked glycosites in industrial strain Aspergillus niger by applying an optimized solid phase enrichment of glycopeptide protocol using hydrazide modified magnetic beads. The enrichment protocol was initially optimized using mouse plasma and A. niger secretome samples, which was then applied to profile N-linked glycosites from both the secretome and whole cell lysates of A. niger. A total of 847 unique N-linked glycosites and 330 N-linked glycoproteins were confidently identified by LC-MS/MS. Based on gene ontology analysis, the identified N-linked glycoproteins in the whole cell lysate were primarily localized in the plasma membrane, endoplasmic reticulum, golgi apparatus, lysosome, and storage vacuoles. The identified N-linked glycoproteins are involved in a wide range of biological processes including gene regulation and signal transduction, protein folding and assembly, protein modification and carbohydrate metabolism. The extensive coverage of glycosylated N-linked glycosites along with identification of partial N-linked glycosylation in those enzymes involving in different biochemical pathways provide useful information for functional studies of N-linked glycosylation and their biotechnological applications in A. niger.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 1035423
• Report Number(s): PNNL-SA-82556; 42901; KP1501021; TRN: US201204%%594
• Journal Information: Journal of Proteome Research, Vol. 11, Issue 1
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
ASPERGILLUS
CARBOHYDRATES
CHEMISTRY
ENDOPLASMIC RETICULUM
ENZYMES
FUNCTIONALS
FUNGI
GENE REGULATION
GENES
GLYCOPROTEINS
GOLGI COMPLEXES
HYDRAZIDES
MASS SPECTROSCOPY
METABOLISM
MODIFICATIONS
NIGER
PLASMA
PROTEINS
STORAGE
STRAINS
filamentous fungi
Aspergillus niger
N-linked glycosylation
glycosites
hydrazide chemistry
secretome.
Environmental Molecular Sciences Laboratory