Difference between revisions of "Part:BBa K5121016"

 
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<partinfo>BBa_K5121016 short</partinfo>
 
<partinfo>BBa_K5121016 short</partinfo>
  
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== Biology ==
  
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RebB encodes an 13 kDa protein which forms one of the main structural components of type 51 R bodies. This part is based off of the wildtype rebB part — BBa_K2912001 — uploaded by SZU-China in 2019, however has been modified to be compatible with cysteine maleimide conjugation at the C-terminus. This is to allow for conjugation with entire R bodies — note that R bodies should be assembled and purified prior to conjugation with the constituent rebB monomers. Our team aimed to use conjugation to attach drugs onto R bodies for use in drug delivery.
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===Usage and Biology===
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Cysteine maleimide conjugation is a form of Michael addition, in which the thiol of the cysteine acts as a nucleophile to react with maleimide, forming a thiosuccinimide adduct. Through this reaction, drugs with maleimide groups can hence be reacted onto proteins with readily accessible cysteines.
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  <figure style="text-align: center;">
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    <img src="https://static.igem.wiki/teams/5121/rebcm/screenshot-2024-09-30-at-12-07-08-am.png" width="80%">
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    <figcaption>
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      Figure 1. Cysteine maleimide conjugation reaction mechanism.
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    </figcaption>
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Since wildtype rebB does not have any cysteines (and nor does RebA), maleimide conjugation provides an ideal conjugation method for orthogonal attachment on to R bodies. This part contains an C-terminal cysteine that has been added to facilitate C-terminal conjugation using cysteine maleimide addition. See the design page for more details.
  
 
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Revision as of 12:28, 2 October 2024


rebB C-terminal CGGGGS

Biology

RebB encodes an 13 kDa protein which forms one of the main structural components of type 51 R bodies. This part is based off of the wildtype rebB part — BBa_K2912001 — uploaded by SZU-China in 2019, however has been modified to be compatible with cysteine maleimide conjugation at the C-terminus. This is to allow for conjugation with entire R bodies — note that R bodies should be assembled and purified prior to conjugation with the constituent rebB monomers. Our team aimed to use conjugation to attach drugs onto R bodies for use in drug delivery.

Cysteine maleimide conjugation is a form of Michael addition, in which the thiol of the cysteine acts as a nucleophile to react with maleimide, forming a thiosuccinimide adduct. Through this reaction, drugs with maleimide groups can hence be reacted onto proteins with readily accessible cysteines.

Figure 1. Cysteine maleimide conjugation reaction mechanism.

Since wildtype rebB does not have any cysteines (and nor does RebA), maleimide conjugation provides an ideal conjugation method for orthogonal attachment on to R bodies. This part contains an C-terminal cysteine that has been added to facilitate C-terminal conjugation using cysteine maleimide addition. See the design page for more details.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]