The relationship between the N-glycosylation of acetylglucosaminyltransferase V and its activity.
OBJECTIVE
The goal of this paper is to investigate the relationship between the N-glycosylation of acetylglucosaminyltransferase V (GlcNAcT-V) and its activity and to know which site among the 6 N-glycosylation sites in the GlcNAcT-V gene is the most important.


METHODS
Wild type of GlcNAcT-V was transfected into COS-7 cells and its activity was measured 48 h later. The first site (Asn 110) was mutated with site-directed mutagenesis and transfected into COS-7 cells.


RESULTS
It was found that after the cells were added tunicamycin (TM, 1 microg/ml), the activity was 11.7% of the wild type. The activity of the cells with mutating GlcNAcT-V was about 12.0% of the wild type. RT-PCR showed that there was no significant change in mRNA expression among the three groups.


CONCLUSIONS
The N-glycosylation is important for its activity. Our results suggest that the N-linked carbohydrates on GlcNAcT-V are required for the posttranscriptional activity of the enzyme.
Zhibing
Z
Z
Shanghai Institute of Biochemistry, Chinese Academy of Sciences, Shanghai 200031.
Huabei
G
G
Chen
W
W
Feng
X
X
Huili
C
C
Youxin
J
J
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D006031:Glycosylation; D016297:Mutagenesis, Site-Directed; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
D012333:RNA, Messenger; D014415:Tunicamycin; D017351:N-Acetylglucosaminyltransferases; C052254:alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase