Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, TbGT15 is present as a single copy per haploid genome, recombinant overexpression of full-length TbGT15 with a C-terminal 3x HA epitope tag, semi-quantitative RT-PCR enzyme expression analysis | Trypanosoma brucei brucei |
Protein Variants | Comment | Organism |
---|---|---|
additional information | gene replacement strategy is used to create TbGT15 null mutant cells and subsequent insertion of tetracycline-inducible ectopic copy. Constructs for gene replacement and ectopic expression are stably transformed into Trypanosoma brucei bloodstream form cells (strain 427, variant 221). Mouse infectivity studies with wild-type and TbGT15 null mutant bloodstream form trypanosomes. No morphological differences between the wild-type and TbGT15 null mutant parasites are ascertained by light microscopy or by scanning electron microscopy. Compared with wild-type cells, the TbGT15 null mutant parasites exhibit slightly slower growth kinetics in vitro, and this mild growth phenotype is partially reversed in TbGT15 conditional null cells grown under permissive conditions. In addition, no difference in its ability to infect mice can be detected for the TbGT15 null mutant. Mutant phenotype, overview | Trypanosoma brucei brucei |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
Golgi membrane | TbGT15 is a type II transmembrane protein and its sequence contains a dibasic motif, which functions as an endoplasmic reticulum exit signal | Trypanosoma brucei brucei | 139 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Trypanosoma brucei brucei |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-GlcNAc + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[VSG221 protein] | Trypanosoma brucei brucei | - |
UDP + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[VSG221 protein] | - |
? | |
UDP-alpha-D-GlcNAc + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[VSG221 protein] | Trypanosoma brucei brucei 927/4 GUTat10.1 | - |
UDP + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[VSG221 protein] | - |
? | |
UDP-GlcNAc + alpha-D-Man-(1->3)-alpha-D-Man-(1->6)-alpha-D-Man | Trypanosoma brucei brucei | - |
? | - |
? | |
UDP-GlcNAc + alpha-D-Man-(1->3)-alpha-D-Man-(1->6)-alpha-D-Man | Trypanosoma brucei brucei 927/4 GUTat10.1 | - |
? | - |
? | |
UDP-GlcNAc + alpha-D-Man-(1->6)-[alpha-D-Man-(1->3)]-beta-D-Man(1->4)-beta-D-GlcNAc-(1->4)-D-GlcNAc | Trypanosoma brucei brucei | - |
UDP + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)-[alpha-D-Man-(1->3)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-D-GlcNAc | - |
? | |
UDP-GlcNAc + alpha-D-Man-(1->6)-[alpha-D-Man-(1->3)]-beta-D-Man(1->4)-beta-D-GlcNAc-(1->4)-D-GlcNAc | Trypanosoma brucei brucei 927/4 GUTat10.1 | - |
UDP + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)-[alpha-D-Man-(1->3)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-D-GlcNAc | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma brucei brucei | Q57U24 | - |
- |
Trypanosoma brucei brucei 927/4 GUTat10.1 | Q57U24 | - |
- |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | enzyme TbGT15 is predominantly expressed in bloodstream form parasites | Trypanosoma brucei brucei | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-GlcNAc + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[VSG221 protein] | - |
Trypanosoma brucei brucei | UDP + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[VSG221 protein] | - |
? | |
UDP-alpha-D-GlcNAc + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[VSG221 protein] | - |
Trypanosoma brucei brucei 927/4 GUTat10.1 | UDP + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[VSG221 protein] | - |
? | |
UDP-GlcNAc + alpha-D-Man-(1->3)-alpha-D-Man-(1->6)-alpha-D-Man | - |
Trypanosoma brucei brucei | ? | - |
? | |
UDP-GlcNAc + alpha-D-Man-(1->3)-alpha-D-Man-(1->6)-alpha-D-Man | - |
Trypanosoma brucei brucei 927/4 GUTat10.1 | ? | - |
? | |
UDP-GlcNAc + alpha-D-Man-(1->6)-[alpha-D-Man-(1->3)]-beta-D-Man(1->4)-beta-D-GlcNAc-(1->4)-D-GlcNAc | - |
Trypanosoma brucei brucei | UDP + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)-[alpha-D-Man-(1->3)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-D-GlcNAc | - |
? | |
UDP-GlcNAc + alpha-D-Man-(1->6)-[alpha-D-Man-(1->3)]-beta-D-Man(1->4)-beta-D-GlcNAc-(1->4)-D-GlcNAc | - |
Trypanosoma brucei brucei 927/4 GUTat10.1 | UDP + beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)-[alpha-D-Man-(1->3)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-D-GlcNAc | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 43100, about, sequence calculation | Trypanosoma brucei brucei |
Synonyms | Comment | Organism |
---|---|---|
GnTII | - |
Trypanosoma brucei brucei |
N-acetylglucosaminyltransferase II | - |
Trypanosoma brucei brucei |
TbGnTII | - |
Trypanosoma brucei brucei |
TbGT15 | - |
Trypanosoma brucei brucei |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Trypanosoma brucei brucei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Trypanosoma brucei brucei |
General Information | Comment | Organism |
---|---|---|
evolution | enzyme TbGT15 belongs to the trypanosome beta3-glycosyltransferase superfamily | Trypanosoma brucei brucei |
malfunction | deletion of TbGT15 is accompanied by the absence of complex N-glycans, as well as alterations in the biosynthesis of the giant poly-Lac-dently from prior TbGnTI action, which is in contrast to canonical GnTIIs that only use substrates after modification by GnTI, i.e. Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc | Trypanosoma brucei brucei |
metabolism | N-acetylglucosaminyltransferase I or II initiate the elaboration of the Manalpha1-3 and Manalpha1-6 arms, respectively, of the conserved trimannosyl-N-acetylchitobiosyl core of N-linked glycans. Proposed scheme for poly-LacNAc-containing N-glycans of bloodstream form trypanosomes, overview | Trypanosoma brucei brucei |
additional information | the enzyme sequence contains a DXD motif, which is generally involved in catalytic activity of known GTs | Trypanosoma brucei brucei |
physiological function | TbGT15 is the glycosyltransferase responsible for the transfer of beta1-2-linked GlcNAc to the alpha1-6-linked alpha-D-mannopyranosyl residue of Manalpha1-6(Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc. The enzyme initiates the elaboration of the Manalpha1-6 arms of the conserved trimannosyl-N-acetylchitobiosyl core of N-linked glycans, structure and biosynthesis of complex N-glycans in the human pathogen are analyzed, and the adaptation by trypanosomes of beta3-glycosyltransferase family members to catalyze beta-1-2 glycosidic linkages is demonstrated. TbGT15 is a non-essential gene in Trypanosoma brucei bloodstream form cells | Trypanosoma brucei brucei |