Probing Protein-Protein Interactions with a Genetically Encoded Photo-crosslinking Amino Acid

• Published in: Chembiochem : a European journal of chemical biology Vol. 12; no. 12; pp. 1854 - 1857
• Main Authors: Ai, Hui-wang, Shen, Weijun, Sagi, Amit, Chen, Peng R., Schultz, Peter G.
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 16.08.2011; WILEY‐VCH Verlag
• Subjects: amber suppression; Amino Acids - chemistry; Amino Acids - genetics; Amino Acids - metabolism; Amino Acyl-tRNA Synthetases - genetics; Amino Acyl-tRNA Synthetases - metabolism; Cross-Linking Reagents - chemistry; Cyclin-Dependent Kinase 5 - chemistry; Cyclin-Dependent Kinase 5 - genetics; Cyclin-Dependent Kinase 5 - metabolism; Diazomethane - metabolism; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Fatty Acids - metabolism; HEK293 Cells; Humans; Lysine - analogs & derivatives; Lysine - metabolism; Molecular Imaging - methods; p21-Activated Kinases - chemistry; p21-Activated Kinases - genetics; p21-Activated Kinases - metabolism; photo-crosslinking; Photochemical Processes - radiation effects; Plasmids; Protein Binding; protein-protein interactions; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RNA, Transfer - genetics; RNA, Transfer - metabolism; Signal Transduction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Transfection; Transformation, Bacterial; Ultraviolet Rays; unnatural amino acids
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.201100194

A pyrrolysyl tRNA/tRNA sythetase pair has been evolved that efficiently incorporates an aliphatic diazirine amino acid into proteins in E. coli and mammalian cells. When site‐specifically introduced into the active site of Cdk5, this unnatural amino acid was able to photo‐crosslink the kinase to its substrate, Pak1.