Difference between revisions of "Part:BBa K5246027"
| Line 28: | Line 28: | ||
DeepTMHMM analysis predicted that HfsJ is a globular protein located on the cytoplasmic side of the membrane. | DeepTMHMM analysis predicted that HfsJ is a globular protein located on the cytoplasmic side of the membrane. | ||
| − | AlphaFold 3 structure, with a high confidence score, shows that HfsJ is most likely a globular protein mostly made of alpha helices. 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. | + | AlphaFold 3 structure, with a high confidence score, shows that HfsJ is most likely a globular protein mostly made of alpha helices. 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). |
Based on our findings and prior research, we propose that HfsJ is likely a globular protein responsible for transferring UDP-N-acetyl-D-mannosaminuronic acid and catalyzing the formation of a glycosidic bond. [1][2] | Based on our findings and prior research, we propose that HfsJ is likely a globular protein responsible for transferring UDP-N-acetyl-D-mannosaminuronic acid and catalyzing the formation of a glycosidic bond. [1][2] | ||
| + | |||
| + | <center> https://static.igem.wiki/teams/5246/registry/hfsj.png </center> | ||
| + | |||
| + | <center> <b> Fig. 1. </b> AlphaFold 3 structure showing </center> | ||
===References=== | ===References=== | ||
Revision as of 14:11, 28 September 2024
CB2/CB2A HfsJ Glycosyltransferase, 6xHis tag for purification
Introduction
Usage and Biology
TBA
This part also has a non his-tagged variant BBa_K5246010.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 632
Illegal BamHI site found at 780 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 353
- 1000COMPATIBLE WITH RFC[1000]
Experimental characterization
Bioinformatic analysis
CDD analysis revealed that HfsJ is part of the WecB/TgA/CpsF glycosyltransferase family. This family catalyzes the formation of glycosidic bonds and may be involved in the biosynthesis of repeating polysaccharide units found in membrane glycolipids. It has domains very similar to E. coli WecG glycosyltransferase, which is responsible for UDP-N-acetyl-D-mannosaminuronic acid transfer. Results are supported by the protein BLAST, which showed significant similarities with the same WecG glycosyltransferase from E. coli.
DeepTMHMM analysis predicted that HfsJ is a globular protein located on the cytoplasmic side of the membrane.
AlphaFold 3 structure, with a high confidence score, shows that HfsJ is most likely a globular protein mostly made of alpha helices. 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).
Based on our findings and prior research, we propose that HfsJ is likely a globular protein responsible for transferring UDP-N-acetyl-D-mannosaminuronic acid and catalyzing the formation of a glycosidic bond. [1][2]
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. Chepkwony, N.K., Berne, C. and Brun, Y.V. (2019b) ‘Comparative analysis of ionic strength tolerance between freshwater and marine Caulobacterales adhesins’, Journal of Bacteriology, 201(18). doi:10.1128/jb.00061-19.