Difference between revisions of "Part:BBa K5246028"
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===Usage and Biology===
 
===Usage and Biology===
 
TBA
 
TBA
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 +
<html>
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<body>
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<p>
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This part also has a non his-tagged variant <a href="https://parts.igem.org/Part:BBa_K5246011">BBa_K5246011</a>.
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</p>
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</html>
  
 
===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K5246028 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5246028 SequenceAndFeatures</partinfo>
  
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===Functional Parameters===
 
−
<partinfo>BBa_K5246028 parameters</partinfo>
 
  
 
===Experimental characterization===
 
===Experimental characterization===
  
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====Bioinformatic analysis====
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Using CDD analysis, it was identified that HfsK is similar to the GNAT N-acetyltransferase family. Its domains suggest that HfsK is part of the Bcls superfamily. Acetyltransferases of this superfamily are usually involved in cellulose biosynthesis. Protein BLAST did not give conclusive results, which could result from a unique HfsK protein amino acid sequence and structure.
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 +
DeepTMHMM's protein topology predictions showed that HfsK is most likely a globular protein located on the cytoplasmic side of the membrane.
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High confidence scores of AlphaFold 3 structures suggest that HfsK is likely a globular protein. 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).
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To summarise, HfsK is most likely a globular N-acetyltransferase. Earlier evidence, combined with our findings, suggests that it plays a role in the deacetylation of N-acetylglucosamine within the holdfast synthesis pathway. [1][2][3]
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<center> https://static.igem.wiki/teams/5246/registry/hfsk.png </center>
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<center> <b> Fig. 1. </b> AlphaFold 3 structure showing </center>
  
 
===References===
 
===References===
 +
1. Chepkwony, N.K. and Brun, Y.V. (2021) ‘A polysaccharide deacetylase enhances bacterial adhesion in high-ionic-strength environments’, iScience, 24(9), p. 103071. doi:10.1016/j.isci.2021.103071.
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<br>
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2. Sprecher, K.S. et al. (2017) ‘Cohesive properties of the Caulobacter crescentus holdfast adhesin are regulated by a novel C-di-GMP effector protein’, mBio, 8(2). doi:10.1128/mbio.00294-17.
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<br>
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3. Hershey, D.M., Fiebig, A. and Crosson, S. (2019) ‘A genome-wide analysis of adhesion in Caulobacter crescentus identifies new regulatory and biosynthetic components for holdfast assembly’, mBio, 10(1). doi:10.1128/mbio.02273-18.

Revision as of 14:21, 28 September 2024


CB2/CB2A HfsK Acetyltransferase, 6xHis tag for purification

Introduction

Usage and Biology

TBA

This part also has a non his-tagged variant BBa_K5246011.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 94
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 94
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 823
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 94
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 94
  • 1000
    COMPATIBLE WITH RFC[1000]


Experimental characterization

Bioinformatic analysis

Using CDD analysis, it was identified that HfsK is similar to the GNAT N-acetyltransferase family. Its domains suggest that HfsK is part of the Bcls superfamily. Acetyltransferases of this superfamily are usually involved in cellulose biosynthesis. Protein BLAST did not give conclusive results, which could result from a unique HfsK protein amino acid sequence and structure.

DeepTMHMM's protein topology predictions showed that HfsK is most likely a globular protein located on the cytoplasmic side of the membrane.

High confidence scores of AlphaFold 3 structures suggest that HfsK is likely a globular protein. 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).

To summarise, HfsK is most likely a globular N-acetyltransferase. Earlier evidence, combined with our findings, suggests that it plays a role in the deacetylation of N-acetylglucosamine within the holdfast synthesis pathway. [1][2][3]

hfsk.png
Fig. 1. AlphaFold 3 structure showing

References

1. Chepkwony, N.K. and Brun, Y.V. (2021) ‘A polysaccharide deacetylase enhances bacterial adhesion in high-ionic-strength environments’, iScience, 24(9), p. 103071. doi:10.1016/j.isci.2021.103071.
2. Sprecher, K.S. et al. (2017) ‘Cohesive properties of the Caulobacter crescentus holdfast adhesin are regulated by a novel C-di-GMP effector protein’, mBio, 8(2). doi:10.1128/mbio.00294-17.
3. Hershey, D.M., Fiebig, A. and Crosson, S. (2019) ‘A genome-wide analysis of adhesion in Caulobacter crescentus identifies new regulatory and biosynthetic components for holdfast assembly’, mBio, 10(1). doi:10.1128/mbio.02273-18.