Coding

Part:BBa_K5246010

Designed by: Edgaras Zaboras   Group: iGEM24_Vilnius-Lithuania   (2024-09-22)
Revision as of 17:57, 27 September 2024 by Zaboras26 (Talk | contribs)


CB2/CB2A HfsJ Glycosyltransferase

Introduction

Usage and Biology

Gene HfsJ from Caulobacter crescentus encodes a protein of 316aa. HfsJ is one of the glycosiltransferases involved in holfast synthesis pathway and is structurally very similar to glycosiltransferases that transfer UDP-N-acetyl-D-mannosaminuronic acid.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 632
    Illegal BamHI site found at 780
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 353
  • 1000
    COMPATIBLE WITH RFC[1000]


Experimental characterization

Bioinformatic analysis

CDD analysis revealed that HfsJ is part of the WecB/TgA/CpsF glycosyltransferase family, which 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, 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]

hfsj.png, hfsa-2.png
Fig. 1. AlphaFold 3 structure showing

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.

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