3.2.1.63 | glycoprotein |
prediction of three N-glycosylation sites on Asn187, Asn280, and Asn401 (residues numbered for the protein without signal peptide). Deglycosylation of the recombinant enzyme expressed from Pichia pastoris by treatment with Endo H or peptide N-glycosidase F. The N-glycans on Asn-280 of the TIM barrel domain retained a chitobiose core and high mannose type substitutions, GlcNAc-beta1-4GlcNAc-beta1-4(Man-alpha1-6)Man-alpha1-3Man-alpha1-2Man-alpha1-2Man, possibly protected from EndoH by the beta,gamma-crystallin domain. The other two N-glycosylation sites Asn187 and Asn401 are each attached to one GlcNAc residue, indicative of Endo H cleavage. No effect of Endo H deglycosylation on the catalytic efficiency with small fucosylated substrates or on thermal stability, but higher solubility of the glycosylated than the deglycosylated enzyme |
732150 |