Difference between revisions of "Part:BBa K4960038"
CirceTerry (Talk | contribs) |
|||
| (17 intermediate revisions by 4 users not shown) | |||
| Line 3: | Line 3: | ||
<partinfo>BBa_K4960038 short</partinfo> | <partinfo>BBa_K4960038 short</partinfo> | ||
| − | |||
| − | + | A flexible linker which can influence the structure of tail fiber connected to the CKGGRAKDC. | |
| + | ===Profile=== | ||
| + | Name: pvc1-pvc12,pvc13_NTD-1*EAAAK-CKGGRAKDC-1*EAAAK -pVc13_CTD,pvc14-pvc16<br> | ||
| + | Origin: Synthetic<br> | ||
| + | Properties: A flexible linker which can influence the structure of tail fiber connected to the CKGGRAKDC.<br> | ||
<!-- Add more about the biology of this part here--> | <!-- Add more about the biology of this part here--> | ||
| + | |||
===Usage and Biology=== | ===Usage and Biology=== | ||
The system is a syringe-like macromolecular complex that inject protein payloads into fat cells by driving a spike through the cellular membrane. Different payloads can be load into the lumen of the inner tube behind the spike, form fusion proteins with the tube, or associate with the spike itself. Studies suggest that the tail fibre is probably involved in target recognition. [1]Our project has enabled the system to target adipocytes by modifying the tail fibre to bind to CKGGRAKDC. CKGGRAKDC is a peptide motif that homes to white fat vasculature. The studies show that CKGGRAKDC targets the white adipose vasculature without a detectable preference for any particular anatomical white fat depot. Meanwhile, the motif is preferentially internalized by a receptor in the adipose vasculature that may serve for targeted delivery of therapeutic compounds to fat.[2] After specific recognition and binding to adipocytes, it can cross the via sheath contraction and inject the desired payloads. | The system is a syringe-like macromolecular complex that inject protein payloads into fat cells by driving a spike through the cellular membrane. Different payloads can be load into the lumen of the inner tube behind the spike, form fusion proteins with the tube, or associate with the spike itself. Studies suggest that the tail fibre is probably involved in target recognition. [1]Our project has enabled the system to target adipocytes by modifying the tail fibre to bind to CKGGRAKDC. CKGGRAKDC is a peptide motif that homes to white fat vasculature. The studies show that CKGGRAKDC targets the white adipose vasculature without a detectable preference for any particular anatomical white fat depot. Meanwhile, the motif is preferentially internalized by a receptor in the adipose vasculature that may serve for targeted delivery of therapeutic compounds to fat.[2] After specific recognition and binding to adipocytes, it can cross the via sheath contraction and inject the desired payloads. | ||
| − | The linker 1*EAAAK on both sides of CKGGRAKDC can affect the function and efficiency of bond and recognition by affecting the structure of the tail protein. | + | The linker 1*EAAAK on both sides of CKGGRAKDC can affect the function and efficiency of bond and recognition by affecting the structure of the tail protein.('''Fig.1''') |
<html> | <html> | ||
<figure class="figure"> | <figure class="figure"> | ||
| − | + | ||
| − | <img src="https://static.igem.wiki/teams/4960/wiki/composite-part/different-linkers.png" class="figure-img img-fluid rounded" height="500px"> | + | <img src="https://static.igem.wiki/teams/4960/wiki/composite-part/different-linkers.png" class="figure-img img-fluid rounded" height="500px"><br> |
| − | + | ||
| − | < | + | |
</figure> | </figure> | ||
</html> | </html> | ||
| − | + | '''Figure 1 AlphaFold2 based prediction of engineered PVC tail fiber trimer structure.''' Structure of adipose-targeting CKGGRAKDC peptide-presenting PVC tail fiber with indicated linkers were shown. <br> | |
| − | + | <html> | |
| + | <figure class="figure"> | ||
| + | <img src="https://static.igem.wiki/teams/4960/wiki/composite-part/bsal-replace/beautiful-pvc.png" class="figure-img img-fluid rounded" height="100px"> | ||
| + | |||
| + | </figure> | ||
| + | |||
| + | </html> | ||
| + | |||
| + | '''Figure 2.PVC composition diagram'''<br> | ||
<!-- Uncomment this to enable Functional Parameter display--> | <!-- Uncomment this to enable Functional Parameter display--> | ||
| + | |||
===Special Design=== | ===Special Design=== | ||
Use the CKGGRAKDC to target fat cells. | Use the CKGGRAKDC to target fat cells. | ||
| Line 38: | Line 49: | ||
<!-- --> | <!-- --> | ||
===Reference=== | ===Reference=== | ||
| − | [1]Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F. Programmable protein delivery with a bacterial contractile injection system. Nature. 2023 Apr;616(7956):357-364 | + | [1]Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F. Programmable protein delivery with a bacterial contractile injection system. Nature. 2023 Apr;616(7956):357-364.<br> |
| − | [2]Kolonin MG, Saha PK, Chan L, Pasqualini R, Arap W. Reversal of obesity by targeted ablation of adipose tissue. Nat Med. 2004 Jun;10(6):625-32 | + | [2]Kolonin MG, Saha PK, Chan L, Pasqualini R, Arap W. Reversal of obesity by targeted ablation of adipose tissue. Nat Med. 2004 Jun;10(6):625-32. |
Latest revision as of 10:14, 12 October 2023
pPVC promoter->pvc1-pvc12->pvc13_NTD-1*EAAAK-CKGGRAKDC-1*EAAAK -pVc13_CTD->pvc14-pvc16
A flexible linker which can influence the structure of tail fiber connected to the CKGGRAKDC.
Profile
Name: pvc1-pvc12,pvc13_NTD-1*EAAAK-CKGGRAKDC-1*EAAAK -pVc13_CTD,pvc14-pvc16
Origin: Synthetic
Properties: A flexible linker which can influence the structure of tail fiber connected to the CKGGRAKDC.
Usage and Biology
The system is a syringe-like macromolecular complex that inject protein payloads into fat cells by driving a spike through the cellular membrane. Different payloads can be load into the lumen of the inner tube behind the spike, form fusion proteins with the tube, or associate with the spike itself. Studies suggest that the tail fibre is probably involved in target recognition. [1]Our project has enabled the system to target adipocytes by modifying the tail fibre to bind to CKGGRAKDC. CKGGRAKDC is a peptide motif that homes to white fat vasculature. The studies show that CKGGRAKDC targets the white adipose vasculature without a detectable preference for any particular anatomical white fat depot. Meanwhile, the motif is preferentially internalized by a receptor in the adipose vasculature that may serve for targeted delivery of therapeutic compounds to fat.[2] After specific recognition and binding to adipocytes, it can cross the via sheath contraction and inject the desired payloads.
The linker 1*EAAAK on both sides of CKGGRAKDC can affect the function and efficiency of bond and recognition by affecting the structure of the tail protein.(Fig.1)
Figure 2.PVC composition diagram
Special Design
Use the CKGGRAKDC to target fat cells. The different linker(1*EAAAK or 3*EAAAK) on both sides of the CKGGRAKDC can affect the structure and function of the tail protein. By altering the linker on both sides of the CKGGRAKDC, we explored the most suitable structure of the tail protein for recognition and bond. At the same time, because of the existence of linker, we can use Golden Gate to replace the sequences in the middle of linker more efficiently than enzyme cutting and enzyme linking.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7406
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2407
Illegal BglII site found at 11264
Illegal BglII site found at 14099
Illegal BglII site found at 16960
Illegal XhoI site found at 2691 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 11792
Illegal NgoMIV site found at 17491
Illegal NgoMIV site found at 17630
Illegal AgeI site found at 6704
Illegal AgeI site found at 9641
Illegal AgeI site found at 10327
Illegal AgeI site found at 10468
Illegal AgeI site found at 11576
Illegal AgeI site found at 17929
Illegal AgeI site found at 19617 - 1000COMPATIBLE WITH RFC[1000]
Reference
[1]Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F. Programmable protein delivery with a bacterial contractile injection system. Nature. 2023 Apr;616(7956):357-364.
[2]Kolonin MG, Saha PK, Chan L, Pasqualini R, Arap W. Reversal of obesity by targeted ablation of adipose tissue. Nat Med. 2004 Jun;10(6):625-32.