Title : Characterization of the N-oligosaccharides attached to the atypical Asn-X-Cys
sequence of recombinant human
epidermal growth factor receptor
Abstract :
- The extracellular domain of human EGF receptor ( sEGFR ) produced by CHO cells has been used in various biophysical studies to elucidate the molecular mechanism of EGF-induced receptor activation
- We have found that the CHO sEGFR contains one oligosaccharide chain attached to an atypical N-glycosylation consensus sequence , Asn(32 )-X( 33 )-Cys(34 )
- The oligosaccharide structure at Asn(32 ) is a mixture of the monosialo and asialo forms of a core fucosylated biantennary complex-type oligosaccharide
- Deletion of this atypical glycosylation site by replacement of Asn(32 ) with lysine changed neither the expression nor function of the full length EGFR in CHO cells
- The glycosylation at Asn(32 ) in CHO sEGFR was incomplete: 20% of Asn(32 ) remained unmodified
- Thus, CHO sEGFR itself is heterogeneous with respect to the glycosylation at Asn(32 ) , which may cause problems in biophysical studies
- An attempt to remove the oligosaccharide at Asn(32 ) enzymatically did not succeed under nondenaturing conditions
- Therefore, sEGFR with the mutation of Asn(32) -> Lys(32 ) is useful for biophysical and biochemical studies, and, particularly, for X-ray crystallography
Output (sent_index, trigger,
protein,
sugar,
site):
- 1. used, , -, -, domain
- 2. attached, , -, one oligosaccharide chain, sequence
- 2. contains, , CHO sEGFR, one oligosaccharide chain, -
- 3. Asn, , -, The oligosaccharide structure, Asn(32 )
- 3. fucosylated, , -, a core fucosylated biantennary complex-type oligosaccharide, -
- 4. glycosylation, , -, -, site
- 5. glycosylation, , -, -, Asn(32 )
- 5. glycosylation, , CHO sEGFR, -, Asn(32 )
- 6. glycosylation, , -, -, Asn(32 )
Output(Part-Of) (sent_index,
protein,
site):
- 0. epidermal growth factor receptor, sequence
- 1. EGF receptor, domain
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):
- 3. epidermal growth factor receptor, The oligosaccharide structure, Asn(32 )
- 5. CHO sEGFR, -, Asn(32 )
- 6. epidermal growth factor receptor, -, Asn(32 )