PMID: 9572875

 

    Legend: Gene, Sites

Title : Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6

Abstract :
  1. The actions of insulin-like growth factors ( IGFs ) are modulated by a family of high-affinity binding proteins ( IGFBPs ), including IGFBP-6 , which preferentially binds IGF-II and is O-glycosylated
  2. Glycosylated and nonglycosylated recombinant human IGFBP-6 , expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography
  3. Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition
  4. Major glycoforms contained 8-16 monosaccharides, including N-acetylhexosamine, hexose, and N-acetylneuraminic acid
  5. Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography
  6. One oligosaccharide chain contained 5-6 monosaccharides, whereas the others contained 2-4 monosaccharides
  7. Glycosylated IGFBP-6 exhibited greater resistance to proteolysis by chymotrypsin and trypsin than nonglycosylated IGFBP-6
  8. Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments , revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected
  9. A model of IGFBP-6 is developed in which these distinct domains are separated by a central region which is O-glycosylated
Output (sent_index, trigger, protein, sugar, site):
  • 0. O-glycosylation, , -, -, sites
  • 0. factor, , insulin-like growth factor binding protein 6, carbohydrate structure, -
  • 2. nonglycosylated, , IGFBP-6, -, -
  • 3. glycosylated, , IGFBP-6, -, -
  • 3. heterogeneity, , -, carbohydrate composition, -
  • 7. nonglycosylated, , IGFBP-6, -, -
  • 9. O-glycosylated, , -, -, region
Output(Part-Of) (sent_index, protein, site):
  • 5. IGFBP-6, Thr126, Ser144, Thr145, Thr146, and Ser152
  • 5. IGFBP-6, sites
  • 8. IGFBP-6, positions
*Output_Site_Fusion* (sent_index, protein, sugar, site):

 

 

Protein NCBI ID SENTENCE INDEX
IGFBPs 3489 1
IGFBP-6 3489 1,2,3,5,7,8,9
Glycosylated IGFBP-6 3489 7
IGF-II 3481 1,2