Difference between revisions of "Part:BBa K5246033"
Line 8: Line 8:
 
===Usage and Biology===
 
===Usage and Biology===
 
TBA
 
TBA
 +
 +
<html>
 +
<body>
 +
<p>
 +
This part also has a non his-tagged variant <a href="https://parts.igem.org/Part:BBa_K5246023">BBa_K5246023</a>.
 +
</p>
 +
</html>
  
 
===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K5246033 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5246033 SequenceAndFeatures</partinfo>
  
−
 
−
===Functional Parameters===
 
−
<partinfo>BBa_K5246033 parameters</partinfo>
 
  
 
===Experimental characterization===
 
===Experimental characterization===
  
 +
====Bioinformatic analysis====
 +
Using CDD analysis, it was identified that HfsK is similar to the GNAT N-acetyltransferase family. Its domains suggest 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.
 +
 +
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]
  
 
===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.
 +
<br>
 +
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.
 +
<br>
 +
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 15:01, 28 September 2024


HB HfsK Acetyltransferase, 6xHis tag for purification

Introduction

Usage and Biology

TBA

This part also has a non his-tagged variant BBa_K5246023.

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 1018
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1068
  • 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 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.

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]

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.