PMID: 1445902

 

    Legend: Gene, Sites

Title : Determination of the glycosylation patterns, disulfide linkages, and protein heterogeneities of baculovirus-expressed mouse interleukin-3 by mass spectrometry

Abstract :
  1. The primary structure of mouse interleukin-3 ( IL-3 ) expressed by recombinant baculovirus-infected silkworm (Bombyx mori) larvae was analyzed by subjecting isolated IL-3 derived peptides to liquid secondary ion mass spectrometry
  2. Two species of IL-3 were isolated from the silkworm hemolymph by reverse-phase high-pressure liquid chromatography
  3. The major component has M(r)20-22 x 10(3) as determined by SDS-PAGE
  4. Liquid secondary ion mass spectrometric analysis was carried out on the reduced tryptic and endopeptidase lysyl-C peptides of glycosylated and deglycosylated IL-3
  5. These studies provided evidence that (1) Asn-16 is heterogeneously glycosylated with four different oligosaccharides, (2) Asn-86 is either nonglycosylated or has attached to it one oligosaccharide, (3) the N-glycosylation sites Asn-44 and Asn-51 are not glycosylated, and (4) there is no O-glycosylation
  6. Liquid secondary ion mass spectrometric analysis of the unreduced tryptic peptides provided evidence for disulfide linkages between Cys-140 and Cys-79 or Cys-80 and between Cys-17 and Cys-79 or Cys-80
  7. In comparison to the major component, a minor IL-3 species (M(r) 17-19 x 10(3) by SDS-PAGE) isolated from the hemolymph showed no difference with respect to the glycosylation pattern or the disulfide linkages, but it was cleaved between Ala-127 and Ser-128 , and only a disulfide linkage between Cys-140 and Cys-79 or Cys-80 held the molecule together
  8. (ABSTRACT TRUNCATED AT 250 WORDS
Output (sent_index, trigger, protein, sugar, site):
  • 0. glycosylation, , interleukin-3, -, -
  • 4. deglycosylated, , IL-3, -, -
  • 4. glycosylated, , IL-3, -, -
  • 5. glycosylated, , -, (1) Asn-16, -
  • 5. glycosylated, , -, Asn-44, -
  • 5. glycosylated, , -, Asn-51, -
  • 5. glycosylated, , -, the N-glycosylation sites Asn-44 and Asn-51, -
  • 5. nonglycosylated, , -, (2) Asn-86, -
Output(Part-Of) (sent_index, protein, site):
  • 4. IL-3, peptides
*Output_Site_Fusion* (sent_index, protein, sugar, site):

 

 

Protein NCBI ID SENTENCE INDEX
interleukin-3 16187 0,1
IL-3 16187 1,2,4,7