Degradation of HaloTag-fused nuclear proteins using bestatin-HaloTag ligand hybrid molecules. 2015

Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
Institute of Molecular and Cellular Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan. m-ishikawa@iam.u-tokyo.ac.jp.

We have developed a protein knockdown technology using hybrid small molecules designed as conjugates of a ligand for the target protein and a ligand for ubiquitin ligase cellular inhibitor of apoptosis protein 1 (cIAP1). However, this technology has several limitations. Here, we report the development of a novel protein knockdown system to address these limitations. In this system, target proteins are fused with HaloTag to provide a common binding site for a degradation inducer. We designed and synthesized small molecules consisting of alkyl chloride as the HaloTag-binding degradation inducer, which binds to HaloTag, linked to BE04 (2), which binds to cIAP1. Using this system, we successfully knocked down HaloTag-fused cAMP responsive element binding protein 1 (HaloTag-CREB1) and HaloTag-fused c-jun (HaloTag-c-jun), which are ligand-unknown nuclear proteins, in living cells. HaloTag-binding degradation inducers can be synthesized easily, and are expected to be useful as biological tools for pan-degradation of HaloTag-fused proteins.

UI MeSH Term Description Entries
D007930 Leucine An essential branched-chain amino acid important for hemoglobin formation. L-Leucine,Leucine, L-Isomer,L-Isomer Leucine,Leucine, L Isomer
D008024 Ligands A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed) Ligand
D009687 Nuclear Proteins Proteins found in the nucleus of a cell. Do not confuse with NUCLEOPROTEINS which are proteins conjugated with nucleic acids, that are not necessarily present in the nucleus. Nucleolar Protein,Nucleolar Proteins,Nuclear Protein,Protein, Nuclear,Protein, Nucleolar,Proteins, Nuclear,Proteins, Nucleolar
D011993 Recombinant Fusion Proteins Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes. Fusion Proteins, Recombinant,Recombinant Chimeric Protein,Recombinant Fusion Protein,Recombinant Hybrid Protein,Chimeric Proteins, Recombinant,Hybrid Proteins, Recombinant,Recombinant Chimeric Proteins,Recombinant Hybrid Proteins,Chimeric Protein, Recombinant,Fusion Protein, Recombinant,Hybrid Protein, Recombinant,Protein, Recombinant Chimeric,Protein, Recombinant Fusion,Protein, Recombinant Hybrid,Proteins, Recombinant Chimeric,Proteins, Recombinant Fusion,Proteins, Recombinant Hybrid
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D006867 Hydrolases Any member of the class of enzymes that catalyze the cleavage of the substrate and the addition of water to the resulting molecules, e.g., ESTERASES, glycosidases (GLYCOSIDE HYDROLASES), lipases, NUCLEOTIDASES, peptidases (PEPTIDE HYDROLASES), and phosphatases (PHOSPHORIC MONOESTER HYDROLASES). EC 3. Hydrolase
D017362 Cyclic AMP Response Element-Binding Protein A protein that has been shown to function as a calcium-regulated transcription factor as well as a substrate for depolarization-activated CALCIUM-CALMODULIN-DEPENDENT PROTEIN KINASES. This protein functions to integrate both calcium and cAMP signals. CREB Protein,DNA-Binding Protein, Cyclic AMP-Responsive,Cyclic AMP-Responsive DNA-Binding Protein,cAMP Response Element-Binding Protein,Cyclic AMP Responsive DNA Binding Protein,DNA Binding Protein, Cyclic AMP Responsive,cAMP Response Element Binding Protein
D048031 JNK Mitogen-Activated Protein Kinases A subgroup of mitogen-activated protein kinases that activate TRANSCRIPTION FACTOR AP-1 via the phosphorylation of C-JUN PROTEINS. They are components of intracellular signaling pathways that regulate CELL PROLIFERATION; APOPTOSIS; and CELL DIFFERENTIATION. jun N-Terminal Kinase,c-jun Amino-Terminal Kinase,c-jun N-Terminal Kinase,jun-NH2-Terminal Kinase,jun-NH2-Terminal Kinases,Amino-Terminal Kinase, c-jun,JNK Mitogen Activated Protein Kinases,Kinase, jun N-Terminal,N-Terminal Kinase, c-jun,N-Terminal Kinase, jun,c jun Amino Terminal Kinase,c jun N Terminal Kinase,jun N Terminal Kinase,jun NH2 Terminal Kinase,jun NH2 Terminal Kinases
D051034 Inhibitor of Apoptosis Proteins A conserved class of proteins that control APOPTOSIS in both VERTEBRATES and INVERTEBRATES. IAP proteins interact with and inhibit CASPASES, and they function as ANTI-APOPTOTIC PROTEINS. The protein class is defined by an approximately 80-amino acid motif called the baculoviral inhibitor of apoptosis repeat; some members have ubiquitin-protein ligase activity. BIRC2 Protein,Baculoviral IAP Repeat Containing Protein 2,Baculoviral IAP Repeat-Containing 2 Protein,Baculoviral Inhibitor Of Apoptosis Repeat Proteins,Cellular Inhibitor of Apoptosis 1 Protein,IAP Protein (Apoptosis),IAP Repeat Containing Proteins,IAP2 Protein,Inhibitor of Apoptosis 2 Protein,Inhibitor-Of-Apoptosis Protein,c-IAP1 Protein,cIAP1 Protein,Apoptosis Proteins Inhibitor,Baculoviral IAP Repeat Containing 2 Protein,Inhibitor Of Apoptosis Protein

Related Publications

Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
July 2016, Bioorganic & medicinal chemistry,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
April 2010, Journal of the American Chemical Society,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
September 2017, Bio-protocol,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
July 2011, Nature chemical biology,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
January 2018, Journal of medicinal chemistry,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
March 2017, Molecular pharmacology,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
August 2015, ACS chemical biology,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
January 2012, Current chemical genomics,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
August 1989, The Journal of biological chemistry,
Shusuke Tomoshige, and Mikihiko Naito, and Yuichi Hashimoto, and Minoru Ishikawa
September 2019, Molecules (Basel, Switzerland),
Copied contents to your clipboard!