RESUMEN
We describe a lanthanide biosensor that responds to CDK4 kinase activity in melanoma cell extracts through a significant and dose dependent increase in luminescence, thanks to sensitization of a DOTA[Tb3+] complex incorporated into a CDK4 substrate peptide by a unique tryptophan residue in an adjacent phosphoaminoacid binding moiety.
Asunto(s)
Complejos de Coordinación/química , Quinasa 4 Dependiente de la Ciclina/análisis , Colorantes Fluorescentes/metabolismo , Compuestos Heterocíclicos con 1 Anillo/química , Péptidos/metabolismo , Aminopiridinas/farmacología , Bencimidazoles/farmacología , Sitios de Unión , Técnicas Biosensibles/métodos , Línea Celular Tumoral , Complejos de Coordinación/síntesis química , Quinasa 4 Dependiente de la Ciclina/antagonistas & inhibidores , Quinasa 4 Dependiente de la Ciclina/metabolismo , Fluorescencia , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/química , Compuestos Heterocíclicos con 1 Anillo/síntesis química , Humanos , Péptidos/síntesis química , Péptidos/química , Fosforilación , Purinas/farmacología , Roscovitina , Terbio/química , Triptófano/químicaRESUMEN
Connexins (Cxs) are integral membrane proteins that form high-conductance plasma membrane channels, allowing communication from cell to cell (via gap junctions) and from cells to the extracellular environment (via hemichannels). Initially described for their role in joining excitable cells (nerve and muscle), gap junctions (GJs) are found between virtually all cells in solid tissues and are essential for functional coordination by enabling the direct transfer of small signalling molecules, metabolites, ions, and electrical signals from cell to cell. Several studies have revealed diverse channel-independent functions of Cxs, which include the control of cell growth and tumourigenicity. Connexin43 (Cx43) is the most widespread Cx in the human body. The myriad roles of Cx43 and its implication in the development of disorders such as cancer, inflammation, osteoarthritis and Alzheimer's disease have given rise to many novel questions. Several RNA- and DNA-binding motifs were predicted in the Cx43 and Cx26 sequences using different computational methods. This review provides insights into new, ground-breaking functions of Cxs, highlighting important areas for future work such as transfer of genetic information through extracellular vesicles. We discuss the implication of potential RNA- and DNA-binding domains in the Cx43 and Cx26 sequences in the cellular communication and control of signalling pathways.
Asunto(s)
Micropartículas Derivadas de Células/metabolismo , Conexina 43/metabolismo , Conexinas/metabolismo , Exosomas/metabolismo , Dominios y Motivos de Interacción de Proteínas , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/patología , Animales , Transporte Biológico , Comunicación Celular , Proliferación Celular , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/metabolismo , Transformación Celular Neoplásica/patología , Conexina 26 , Conexina 43/genética , Conexinas/genética , Uniones Comunicantes , Humanos , Inflamación , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patología , Osteoartritis/genética , Osteoartritis/metabolismo , Osteoartritis/patología , ARN/genética , ARN/metabolismoRESUMEN
We report a light-sensitive histidine building block for Fmoc/tBu solid-phase peptide synthesis in which the imidazole side chain is coordinated to a ruthenium complex. We have applied this building block for the synthesis of caged-histidine peptides that can be readily deprotected by irradiation with visible light, and demonstrated the application of this approach for the photocontrol of the activity of Ni(II)-dependent peptide nucleases.
Asunto(s)
2,2'-Dipiridil/análogos & derivados , Histidina/química , Luz , Compuestos Organometálicos/química , Péptidos/química , Péptidos/síntesis química , Fotólisis/efectos de la radiación , 2,2'-Dipiridil/química , Estructura Molecular , Péptidos/efectos de la radiaciónRESUMEN
A new 4-(N,N-dimethylamino) phthalimide-based environment-sensitive fluorescent building block for solid phase peptide synthesis, has been synthesized and incorporated into peptides. Peptides incorporating this residue show great potential for biological applications in sensing protein/protein interactions.