Difference between revisions of "Part:BBa K2794002"

 
Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2794002 short</partinfo>
 
<partinfo>BBa_K2794002 short</partinfo>
 +
{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
 +
! colspan="2" style="background:#A31F34;"|<span style="color:#ffffff">Nedd4 Family Interacting Protein 1</span>
 +
|-
 +
|'''Function'''
 +
|Exosome Cargo Active Loading
 +
|-
 +
|'''Use in'''
 +
|Mammalian cells
 +
|-
 +
|'''RFC standard'''
 +
|RFC 10 compatible
 +
|-
 +
|'''Backbone'''
 +
|pSB1C3<br>
 +
|-
 +
|'''Submitted by'''
 +
|[http://2018.igem.org/Team:SHSU_China SHSU_China]
 +
|}
  
−
Nedd4 Family Interacting Protein1
+
==Overview==
 +
Nedd4 Family Interacting Protein 1 (Nedd4, synonym N4WBP5) is a membrane protein located on the Golgi apparatus and endosome. It's a evolutionarily conserved protein with three transmembrane domains. This protein is thought to be part of a family of integral Golgi membrane proteins.
 +
 
 +
As said in the name of the protein, ndfip1 interacts with protein Nedd4, a ubiquitin-protein ligase. Nedd4 has 3-4 WW domain, a carboxyl-terminal Hect (homologous to E6-APCarboxyl terminus) domain and in most cases an amino-terminal C2 domain. Ndfip1 binds Nedd4 WW domains via the two PPXY motifs present in the amino terminus of the protein. 
 +
 
 +
Ndfip1 act as an adaptor for Nedd4-like proteins and their putative targets to control ubiquitin-dependent protein sorting and trafficking, ESCRT pathways that involve sorting ubiquitinated cargo into endosome is majorly involved in transporting protein out of cell using exosomes. The process can be separated to the following steps:
 +
 
 +
1: E3 ubiquitin ligase Nedd4 binds its WW domain to the PPXY motif on Ndfip1, and the cargo that also has ww domain binds to another PPXY motif on Ndfip1.
 +
2: Nedd4 act as scaffolding proteins to indirectly facilitate the transfer of ubiquitin by bringing E2 and substrate proteins into close proximity.
 +
3: The ubiquitinated cargo is loaded into the endosome by the ubiquitination related endosomal sorting pathway ESCRT.
 +
 
 +
[[File:T--SHSU_China--ndfipww.png|400px|]]
 +
 
 +
Hence, co-expression of Ndfip1 and WW tag labeled protein can actively ubiquitinate target protein and made it loaded into exosomes and create exosomes for medical use. This part should be used along with BBa_K2794003 WW Tag3 and BBa_K2794004 WW Tag4.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:49, 13 October 2018


Nedd4 Family Interacting Protein1 (Ndfip1)

Nedd4 Family Interacting Protein 1
Function Exosome Cargo Active Loading
Use in Mammalian cells
RFC standard RFC 10 compatible
Backbone pSB1C3
Submitted by [http://2018.igem.org/Team:SHSU_China SHSU_China]

Overview

Nedd4 Family Interacting Protein 1 (Nedd4, synonym N4WBP5) is a membrane protein located on the Golgi apparatus and endosome. It's a evolutionarily conserved protein with three transmembrane domains. This protein is thought to be part of a family of integral Golgi membrane proteins.

As said in the name of the protein, ndfip1 interacts with protein Nedd4, a ubiquitin-protein ligase. Nedd4 has 3-4 WW domain, a carboxyl-terminal Hect (homologous to E6-APCarboxyl terminus) domain and in most cases an amino-terminal C2 domain. Ndfip1 binds Nedd4 WW domains via the two PPXY motifs present in the amino terminus of the protein.

Ndfip1 act as an adaptor for Nedd4-like proteins and their putative targets to control ubiquitin-dependent protein sorting and trafficking, ESCRT pathways that involve sorting ubiquitinated cargo into endosome is majorly involved in transporting protein out of cell using exosomes. The process can be separated to the following steps:

1: E3 ubiquitin ligase Nedd4 binds its WW domain to the PPXY motif on Ndfip1, and the cargo that also has ww domain binds to another PPXY motif on Ndfip1. 2: Nedd4 act as scaffolding proteins to indirectly facilitate the transfer of ubiquitin by bringing E2 and substrate proteins into close proximity. 3: The ubiquitinated cargo is loaded into the endosome by the ubiquitination related endosomal sorting pathway ESCRT.

T--SHSU China--ndfipww.png

Hence, co-expression of Ndfip1 and WW tag labeled protein can actively ubiquitinate target protein and made it loaded into exosomes and create exosomes for medical use. This part should be used along with BBa_K2794003 WW Tag3 and BBa_K2794004 WW Tag4.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 479
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 36
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 460