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Reverse Dimerization System

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제조사 제품코드 제품명 용량 가격
(부가세별도)		 비고 사용자매뉴얼
Clontech
 635066
 iDimerize Reverse Dimerization System
관련학술 구매하기 라이선스  		each
 2,158,000원  제조사 페이지로 바로가기 x32734, x33559, x33560, x99924, x33561
Clontech
 635053
 D/D Solubilizer
관련학술 구매하기 		5 X 500 ㎕ (635054 x 5)
 2,570,000원  제조사 페이지로 바로가기 x32734
Clontech
 635054
 D/D Solubilizer
관련학술 구매하기 		500 ㎕
 571,000원  제조사 페이지로 바로가기

The iDimerize Reverse Dimerization System brings the disruption of protein complexes under real-time, small molecule control. A protein of interest is fused to the DmrD binding domain, and the fusion protein molecules aggregate unless the D/D Solubilizer ligand is present.

Reverse Dimerization: Disrupting Protein-Protein Interactions

The iDimerize Reverse Dimerization System is a "reverse dimerization" system-aggregation is the resting state, and the D/D Solubilizer breaks up protein-protein interactions. Therefore, the iDimerize Reverse Dimerization System complements inducible dimerization, and can be used in analogous ways to create inducible alleles. In principle, most processes that can be brought under dimerizer control can also be controlled in the reverse manner using this kit to turn off a process that is activated by oligomerization.



Figure 1. Mechanism of the iDimerize Reverse Dimerization System. The Reverse Dimerization System incorporates a binding motif (purple) that causes protein aggregation and a dimerizer (yellow) which can be used to disaggregate (solubilize) the proteins. This system can be used to study intracellular transport and to induce regulated secretion.
Inducible Secretion
The ability to create large protein aggregates has unique applications. For example, adding a secretory signal sequence to fusion proteins allows them to be reversibly stored as aggregates in the endoplasmic reticulum. The ligand can then be added to induce a rapid pulse of protein secretion from the cells. This method has been used to induce rapid, transient and tightly-regulated secretion of human growth hormone (hGH) and insulin (1).

Protein aggregates can also be used in protein trafficking research. For example, this approach has been used to discover the existence of "mega-vesicles" transporting cargo across the Golgi stack (2).



Figure 2. The iDimerize Reverse Dimerization System enables dose-dependent control of protein secretion. Fusion proteins containing DmrD domains localize to the endoplasmic reticulum as massive aggregates (left). When the D/D Solubilizer is added, it dissolves the aggregates and allows the protein to be exported through the secretory apparatus (right). To ensure secretion of the authentic protein, a furin cleavage site is positioned between the DmrD domains and the protein of interest. Since furin is exclusively expressed in the trans Golgi, the fusion protein will be processed as it traverses this compartment, resulting in the secretion of the correctly processed protein.
D/D Solubilizer Ligand
The D/D Solubilizer is a synthetic, cell-permeable ligand that can be used to disrupt dimerization of fusion proteins containing the DmrD domain. The D/D Homodimerizer has been tested in vitro and in mice. It is nontoxic. We suggest testing various D/D Homodimerizer concentrations within the recommended range (10-500 nM) for different lengths of time (30 minutes to 12+ hours) in order to obtain a complete dose-response profile. The D/D Solubilizer performs the same function as the AP21998 ligand, which was previously supplied by ARIAD Pharmaceuticals Inc. It is a different molecule than AP21998.
Applications
  • Rapid, reversible changes in the subcellular location, aggregation state and/or biological activity of engineered proteins, in vitro or in vivo
  • Rapid induction of protein secretion
  • Protein trafficking studies
  • Inducible animal models and cell lines
References
1. Rivera, V. M., et al. (2000) Science 287(5454):826-830. Regulation of protein secretion through controlled aggregation in the endoplasmic reticulum.
2. Volchuk, A., et al. (2000) Cell 102(3):335-348. Megavesicles implicated in the rapid transport of intracisternal aggregates across the Golgi stack.


Keyword : RPD Regulated Secretion/Aggregation Kit