Journal: Molecular & Cellular Proteomics : MCP
Article Title: Unusual N-glycan Structures Required for Trafficking Toxoplasma gondii GAP50 to the Inner Membrane Complex Regulate Host Cell Entry Through Parasite Motility *
Figure Lengend Snippet: Deciphering N -glycan structures and positioning on GAP50 protein. A , GAP50 glycopeptides were identified using nanoLC-MS/MS. The MS/MS spectrum from the parent ion m/z = 1150.48 corresponds to the chymotryptic peptide 227 SNVTSRAW 234 with the N -glycosylation Hex 6 (HexNAc) 2 in position Asn228. B , The MS/MS spectrum from parent ion at m/z = 1329.03 corresponds to the tryptic peptide 136 NYTSEALR 143 with the N -glycosylation Hex 8 (HexNAc) 2 in position Asn136. C , The MS/MS spectrum from parent ion at m/z = 1414.31 corresponds to the tryptic peptide 77 VAANEHISFIASPGSNFLGGVSSLNDTR 104 with the N -glycosylation Hex 6 (HexNAc) 2 in position Asn101. Symbols were as follows: p , peptide; ■, N-acetylglucosamine; ●, mannose; ●, hexose. (D-G) Chymotryptic digestion of GAP50-HAFLAG profile using nanoLC-MS. D , Extracted ion chromatograms ( m/z 1150.0, 1231.0 for peak 1; 1245.5, 1326.5, 1407.5 for peak 2; 1261.0, 1342.1, 1423.1, for peak 3; ± 0.2 Da) indicated the elution zone of glycopeptides assigned by MS/MS. E , Combined mass spectrum of the glycopeptides containing Asn228 eluting at 17 min (peak 1). F , Combined mass spectrum of the glycopeptides containing Asn136 eluting at 19 min (peak 2). G , Combined mass spectrum of the glycopeptides containing Asn101 eluting at 23 min (peak 3).
Article Snippet: Five microliters of chymotrypsin GAP50 digest were analyzed by nanoLC-MS using the nanoACQUITY Ultra-Performance-LC system (UPLC, Waters, Milford, MA) coupled to a SYNAPT High Definition Mass Spectrometry quadrupole time-of-flight tandem mass spectrometer (Waters, Milford, MA) equipped with a nano-electrospray source.
Techniques: Mass Spectrometry