Characterization and analysis of a novel glycoprotein from snake venom using liquid chromatography-electrospray mass spectrometry and Edman degradation.
An N-linked glycosylation in a novel C-lectin protein from snake venom was observed by Edman degradation and liquid chromatography-electrospray mass spectrometry. The peptides obtained by trypsin cleavage were analyzed to confirm the amino acid sequence and Asn5 was found to be the N-glycosylation site. The result was further confirmed by N-glycosidase digestion. In addition, the protein and tryptic peptides with and without glycan chain were characterized by mass spectrometry according to the mass difference. The glycopeptide obtained from proteolytic digestion was analyzed and the glycoforms were identified as high-mannose type by tandem MS coupled with alpha-mannosidase digestion. An oxidized Met residue was detected and located in the protein by mass spectrometry.
Zeng
R
R
Shanghai Institute of Biochemistry, Chinese Academy of Sciences, China.
Xu
Q
Q
Shao
X X
XX
Wang
K Y
KY
Xia
Q C
QC
D000581:Amidohydrolases / Q000378:metabolism; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*; D006031:Glycosylation; D037181:Lectins, C-Type / Q000737:chemistry* / Q000302:isolation & purification*; D008361:Mannosidases / Q000494:pharmacology; D013058:Mass Spectrometry; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D017434:Protein Structure, Tertiary; D012910:Snake Venoms / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013997:Time Factors; D017841:Trimeresurus / Q000378:metabolism; D014357:Trypsin / Q000494:pharmacology; D014757:Viper Venoms / Q000737:chemistry* / Q000302:isolation & purification*; D043323:alpha-Mannosidase
D000596:Amino Acids; D006023:Glycoproteins; D037181:Lectins, C-Type; D012910:Snake Venoms; C468568:Trimeresurus stejnegeri lectin; D014757:Viper Venoms; D008361:Mannosidases; D043323:alpha-Mannosidase; D014357:Trypsin; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase