| Products/Services Used | Details | Operation |
|---|---|---|
| Mutagenesis Services | The DNA, containing at the 5′ end a recognition sequence for XhoI and a KEX2 cleavage signal, and at the 3′ end a sequence encoding a histidine tag, a stop codon and a recognition sequence for SacII, was cloned into the pUC57 vector (GenScript). The DNA sequences encoding the GalNAc-T4 chimeras were also synthesized by GenScript and cloned in the same above vector. The DNA sequences encoding the GalNAc-T2 chimeras were synthesized by GenScript and cloned in the vector pPICZαA vector as previously described12. Site-directed mutagenesis experiments were also performed by GenScript using the previous reported template pPICZαAgalnact2 (K75-Q571)12 and the plasmid encoding the chimera 2. | Get A Quote |
The polypeptide GalNAc-transferases (GalNAc-Ts)， that initiate mucin-type O-glycosylation， consist of a catalytic and a lectin domain connected by a flexible linker. In addition to recognizing polypeptide sequence， the GalNAc-Ts exhibit unique long-range N- and/or C-terminal prior glycosylation (GalNAc-O-Ser/Thr) preferences modulated by the lectin domain. Here we report studies on GalNAc-T4 that reveal the origins of its unique N-terminal long-range glycopeptide specificity， which is the opposite of GalNAc-T2. The GalNAc-T4 structure bound to a monoglycopeptide shows that the GalNAc-binding site of its lectin domain is rotated relative to the homologous GalNAc-T2 structure， explaining their different... More
