Title : Posttranslational N-glycosylation takes place during the normal processing of human
coagulation factor VII
Abstract :
- N-glycosylation is normally a cotranslational process that occurs during translocation of the nascent protein to the endoplasmic reticulum
- In the present study, however, we demonstrate posttranslational N-glycosylation of recombinant human coagulation factor VII ( FVII ) in CHO-K1 and 293A cells
- Human FVII has two N-glycosylation sites ( N145 and N322 )
- Pulse-chase labeled intracellular FVII migrated as two bands corresponding to FVII with one and two N-glycans, respectively
- N-glycosidase treatment converted both of these band into a single band, which comigrated with mutated FVII without N-glycans
- Immediately after pulse, most labeled intracellular FVII had one N-glycan, but during a 1-h chase, the vast majority was processed into FVII with two N-glycans, demonstrating posttranslational N-glycosylation of FVII
- Pulse-chase analysis of N-glycosylation site knockout mutants demonstrated cotranslational glycosylation of N145 but primarily or exclusively posttranslational glycosylation of N322
- The posttranslational N-glycosylation appeared to take place in the same time frame as the folding of nascent FVII into a secretion-competent conformation, indicating a link between the two processes
- We propose that the cotranslational conformation(s) of FVII are unfavorable for glycosylation at N332 , whereas a more favorable conformation is obtained during the posttranslational folding
- This is the first documentation of posttranslational N-glycosylation of a non-modified protein in mammalian cells with an intact N-glycosylation machinery
- Thus, the present study demonstrates that posttranslational N-glycosylation can be a part of the normal processing of glycoproteins
Output (sent_index, trigger,
protein,
sugar,
site):
- 10. N-glycosylation, , protein, -, -
- 11. glycoproteins, , glycoproteins, -, -
- 2. N-glycosylation, , FVII, -, -
- 2. N-glycosylation, , coagulation factor VII, -, -
- 3. N-glycosylation, , -, -, N145 and N322
- 3. N-glycosylation, , -, -, sites
- 3. sites, , -, -, N145 and N322
- 3. sites, , -, -, sites
- 4. FVII, , FVII, one and two N-glycans, -
- 6. FVII, , FVII, two N-glycans, -
- 6. N-glycosylation, , FVII, -, -
- 6. had, , FVII, one N-glycan, -
- 7. N-glycosylation, , -, -, site
- 7. glycosylation, , -, -, N145
- 7. glycosylation, , -, -, N322
- 9. glycosylation, , -, -, N332
Output(Part-Of) (sent_index,
protein,
site):
- 3. Human FVII, N145 and N322
- 3. Human FVII, sites
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):
- 3. Human FVII, -, N145 and N322
- 7. coagulation factor VII, -, N145
- 7. coagulation factor VII, -, N322
- 9. coagulation factor VII, -, N332