RPD Regulated Secretion/Aggregation

2006

• Kwok, C., Zeisig, B.B., Dong, S., and So, C.W. (2006) Forced homo-oligomerization of RARalpha leads to transformation of primary hematopoietic cells. Cancer Cell 9:95-108.
  The RPD system is used to demonstrate that dimerization is required for the transforming activity of RARalpha. Gel filtration was used to confirm the oligomerized state of RARalpha-F36M fusion proteins, and addition of AP21998 was shown to reverse dimerization and inhibit transformation.
• Sawyer, G.W., Ehlert, F.J., and Hart, J.P. (2006) Determination of the rate of muscarinic M1 receptor plasma membrane delivery using a regulated secretion/aggregation system. J Pharmacol Toxicol Methods 53:219-233.
  Uses the RPD system to control the delivery of the human muscarinic M1 receptor, a 7-transmembrane receptor, to the plasma membrane. Timed release of aggregated receptor with AP21998 was used to determine the rate of delivery.

2005

• Brenzel, S., and Mootz, H.D. (2005) Design of an intein that can be inhibited with a small molecule ligand. J Am Chem Soc 127:4176-4177.
  Uses the regulated aggregation system to create an intein that can be inactivated by the monomeric ligand AP21998.
• DeRocher, A., Gilbert, B., Feagin, J.E., and Parsons, M. (2005) Dissection of brefeldin A-sensitive and -insensitive steps in apicoplast protein targeting. J Cell Sci 118:565-574.
  Uses the conditional aggregation system to study protein trafficking in the pathogen Toxoplasma gondii.

2004

• Hansen, J.L., Theilade, J., Haunso, S., and Sheikh, S.P. (2004) Oligomerization of wild type and nonfunctional mutant angiotensin II type I receptors inhibits galphaq protein signaling but not ERK activation. J Biol Chem 279:24108-24115.
  The regulated aggregation system is used to demonstrate that oligomerization of the 7 transmembrane angiotensin II type 1 receptor plays an important role in the activation of downstream signaling pathways.

2003

• Rosen, H., Gao, Y., Johnsson, E., and Olsson, I. (2003) Artificially controlled aggregation of proteins and targeting in hematopoietic cells. J Leukoc Biol 74:800-9.
  Studies the localization of fusion protein aggregates formed using the homodimerization system (FKBP fusion proteins and homodimerizer AP20187) or the RPD system (Fm fusion proteins, disaggregated by AP21998). Concludes that aggregation is not sufficient to induce protein sorting into lysosomes in hematopoietic cells.

2000

• Rivera, V.M., Wang, X., Wardwell, S., Courage, N.L., Volchuk, A., Keenan, T., Holt, D.A., Gilman, M., Orci, L., Cerasoli, F., Jr., Rothman, J.E., and Clackson, T. (2000) Regulation of protein secretion through controlled aggregation in the endoplasmic reticulum. Science 287:826-30.
  Describes a new method for controlling the production of secreted proteins. Proteins are engineered to accumulate as aggregates in the ER, from where they can be rapidly released for processing and secretion by adding ligand. Rapid and transient secretion of growth hormone and insulin is demonstrated in vitro and in vivo, with regulated insulin secretion resulting in a transient correction of serum glucose concentrations in a mouse model of hyperglycemia.
• Rollins, C.T., Rivera, V.M., Woolfson, D.N., Keenan, T., Hatada, M., Adams, S.E., Andrade, L.J., Yaeger, D., van Schravendijk, M.R., Holt, D.A., Gilman, M., and Clackson, T. (2000) A ligand-reversible dimerization system for controlling protein-protein interactions. Proc Natl Acad Sci USA 97:7096-101.
  Details the biochemical basis for the regulated aggregation system, and its uses to control transcription and fusion protein aggregation. The discovery is described of a point mutant of FKBP which homodimerizes in a ligand-reversible manner. The mutant FKBP and ligand form the basis of a reversible aggregation system that can be used to control the activity and secretion of proteins.
• Volchuk, A., Amherdt, M., Ravazzola, M., Brugger, B., Rivera, V.M., Clackson, T., Perrelet, A., Sollner, T.H., Rothman, J.E., and Orci, L. (2000) Megavesicles implicated in the rapid transport of intracisternal aggregates across the Golgi stack. Cell 102:335-48.
  Uses controlled protein aggregation to reveal the existence of megavesicles that can transport large protein aggregates through the Golgi.
• Wandless, T.J. (2000) A confederacy of bunches: fundamentals and applications of a self- associating protein. Proc Natl Acad Sci USA 97:6921-3.
  A minireview on the original papers (Rivera et al., 2000 and Rollins et al., 2000) describing the regulated secretion/aggregation system.