biochemical and molecular analyses of natural products
	Craig Crews, Ph.D. Associate Professor of Molecular, Cellular, & Developmental Biology Room: KBT 400 Phone: (203) 432-9364 / 6553 Email: Craig.Crews@yale.edu Web site B.A. University of Virginia; Ph.D. Harvard University

Targeting Intracellular Proteins for Degradation using Small Molecules. Hela cells (top left) expressing Green Fluorescent Protein fused to the androgen receptor (AR) were incubated with 25 µM DHT-ProTac (above center), which targeted the AR fusion protein for degradation within 60 minutes (top right).

Ectopic Limb Regeneration. A 14 day hind limb blastema from a pigment axolotl (Ambystoma mexicanum) grafted onto the upper leg of an albino axolotl.

Our laboratory is interested in developing novel reagents, which will allow us to explore new areas in cell biology. This 'chemical genetic' approach uses biologically active natural products to identify critical components in intracellular processes. In the past few years, our efforts have focused on three classes of natural products: anti-angiogenic, antitumor, and anti-inflammatory compounds. Using affinity chromatography, we showed that the anti-angiogenic compound fumagillin covalently binds and inhibits the metalloprotease methionine aminopeptidase 2, resulting in endothelial cell-specific cell cycle arrest. A similar approach using the antitumor agent epoxomicin and the anti-inflammatory compound parthenolide revealed that these natural products potently target the 20S proteasome and the IkB kinase IKKb, respectively. In addition to the study of other biologically active natural products, we are pursuing approaches that will allow for the selective degradation of any desired intracellular protein.

A more recent research interest focuses on the molecular mechanisms of limb regeneration. Using the axolotl as a model system, we would like to understand the signals that induce cellular dedifferentiation and regeneration.

Selected Publications

Myung, J., Kim, K., Lindsten, K.K., Dantuma, N.P., and C.M. Crews. (2001) Lack of Proteasome Active Site Allostery as Revealed by Subunit-Specific Inhibitors. Molecular Cell, 7(2):411-420.

Yeh, J. and C.M. Crews (2003) Chemical Genetics: Adding to the Developmental Biology Toolbox Developmental Cell 5(1):11-19

Brdlik, C., and C.M. Crews (2004) A Single Amino Acid Residue Defines Inhibitor Specificity for the Methionine Aminopeptidase Family. J.Biol.Chem. 279:9475-80.

Schneekloth, Jr., J.S., F. Fonseca, M. Koldobskiy, A. Mandal, R. Deshaies, K. Sakamoto, and C. M. Crews (2004) Chemical Genetic Control of Protein Levels: Selective in vivo Targeted Degradation JACS 126(12):3748-54.

top