Part:BBa_K771002

Membrabe Anchor 2:ssDsbA-Lgt-SH3 Domain-MS2

Membrane anchor 2:RNA aptamer binding protein MS2 (BBa_K771112) fused with membrane protein Lgt(BBa_K771102).

Overview: Membrane Scaffold System

Demonstration of the whole Membrane Scaffold device. Membrane Anchor 2, 3, 4, 5 consitutively aggregate while Membrane Anchor 1 and 6 only aggregate with constitutive cluster (Membrane Anchor 2, 3, 4, 5) when certain signal is present.

Backgroud

Signal Induced Aggregation

Membrane Anchor 1 and 2 act as a controlling point to determine whether certain pathway （in this case involving enzyme 1, 2, 3, 4 and 5) is activated or not. RNA molecule D0(BBa_K771009) with PP7 and MS2 aptamer domain acts as the signal.

Sketch of signal RNA molecule (called D0) which consists of PP7 and MS2 aptamer domains. B: Through fusing PP7 and MS2 RNA biding domain (BBa_K771111 BBa_K771112) to our membrane device, RNA molecule D0(BBa_K771009) in Figure A can function as a bridge to connect different proteins, thus decreasing distance between corresponding enzymes.

Constitutive Aggregation

Membrane Anchor 2 and 3 constitutively aggregate through SH3 domain(BBa_K771107) and SH3 ligand (BBa_K771108).

Design

RNA aptamer binding protein MS2(BBa_K771112) fused with membrane protein Lgt (BBa_K771102). When RNA molecule D0 is present, Membrane Anchor 1(BBa_K771001) is expected to dimerize with Membrane Anchor 2 through MS2(BBa_K771112) and PP7(BBa_K771111) domain.

Related Biobrick

<br\>Membrabe Anchor 1([BBa_K771001]) <br\>Membrabe Anchor 3([BBa_K771003]) <br\>Membrabe Anchor 4([BBa_K771004]) <br\>Membrabe Anchor 5([BBa_K771005]) <br\>Membrabe Anchor 6([BBa_K771006])

Reference

Delebecque, C. J., A. B. Lindner, et al. (2011). "Organization of intracellular reactions with rationally designed RNA assemblies." Science 333(6041): 470.

Dueber, J. E., G. C. Wu, et al. (2009). "Synthetic protein scaffolds provide modular control over metabolic flux." Nature biotechnology 27(8): 753-759.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 883
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 504
  Illegal BsaI site found at 1609
  Illegal SapI.rc site found at 1150