Difference between revisions of "Part:BBa K5246007"
| Line 8: | Line 8: | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
HfsG gene encodes a cytoplasmic protein of 309 aa homologous to family 2 glycosyltransferases. Transferrs sugar units from UDP-GlcNAc to oligosaccharide, catalyzes the polymerization of GlcNAc. C.Crescentus HfsG mutants were completely devoid of holdfast material | HfsG gene encodes a cytoplasmic protein of 309 aa homologous to family 2 glycosyltransferases. Transferrs sugar units from UDP-GlcNAc to oligosaccharide, catalyzes the polymerization of GlcNAc. C.Crescentus HfsG mutants were completely devoid of holdfast material | ||
| + | |||
| + | <html> | ||
| + | <body> | ||
| + | <p> | ||
| + | This part also has a non his-tagged variant <a href="https://parts.igem.org/Part:BBa_K5246025">BBa_K5246025</a>. | ||
| + | </p> | ||
| + | </html> | ||
===Sequence and Features=== | ===Sequence and Features=== | ||
Revision as of 12:49, 28 September 2024
CB2/CB2A HfsG Glycosyltransferase
Introduction
Usage and Biology
HfsG gene encodes a cytoplasmic protein of 309 aa homologous to family 2 glycosyltransferases. Transferrs sugar units from UDP-GlcNAc to oligosaccharide, catalyzes the polymerization of GlcNAc. C.Crescentus HfsG mutants were completely devoid of holdfast material
This part also has a non his-tagged variant BBa_K5246025.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 825
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 28
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 480
- 1000COMPATIBLE WITH RFC[1000]
Experimental characterization
Bioinformatic analysis
CDD and protein BLAST analysis suggest that HfsG is a glucosyltransferase family 2 protein. Proteins of this family are involved in cell wall biosynthesis. HfsG is similar to the WecA protein in E.Coli that catalyzes the transfer of the GlcNAc-1-phosphate moiety from UDP-GlcNAc onto the carrier lipid undecaprenyl phosphate. In the case of HfsG, it catalyzes the transfer of UDP-GlcNAc to the sugar acceptor made earlier in the holdfast synthesis pathway.
Protein topology analysis using DeepTMHMM suggests that HfsG is a globular protein located in the cytoplasm. AlphaFold 3 structures further confirm it. A pTM score above 0.5 suggests that the predicted overall structure may closely resemble the true protein fold, while ipTM indicates the accuracy of the subunit positioning within the complex. Values higher than 0.8 represent confident, high-quality predictions (Fig. 1).
HfsG is family 2 glycosyltransferase similar to WecA of E.Coli. This globular protein transfers UDP-GlcNAc to the acceptor molecule, our conclusions are in agreement with existing research. [1][2][3]
References
1. Toh, E., Kurtz, Harry D. and Brun, Y.V. (2008) ‘Characterization of the Caulobacter crescentus holdfast polysaccharide biosynthesis pathway reveals significant redundancy in the initiating glycosyltransferase and polymerase steps’, Journal of Bacteriology, 190(21), pp. 7219–7231. doi:10.1128/jb.01003-08.
2. Hardy, G.G. et al. (2018) ‘Mutations in sugar-nucleotide synthesis genes restore holdfast polysaccharide anchoring to Caulobacter crescentus holdfast anchor mutants’, Journal of Bacteriology, 200(3). doi:10.1128/jb.00597-17.
3. Sulkowski, N.I. et al. (2019) ‘A multiprotein complex anchors adhesive holdfast at the outer membrane of Caulobacter crescentus’, Journal of Bacteriology, 201(18). doi:10.1128/jb.00112-19.