A photoregulated ligand for the nuclear import receptor karyopherin alpha.
The ability to orchestrate the transport of proteins between nucleus and cytoplasm provides cells with a powerful regulatory mechanism. Selective translocation between these compartments is often used to propagate cellular signals, and it is an intimate part of the processes that control cell division, viral replication, and other cellular events. Therefore, precise experimental control over protein localization, through the agency of light, would provide a powerful tool for the study and manipulation of these events. To this end, a prototype photoregulated nuclear localization signal (NLS) was derived from a native NLS. A library of 30 mutants of the bipartite NLS from Xenopus laevis nucleoplasmin containing a novel, photoisomerizable amino acid was prepared by parallel, solid-phase synthesis and screened in vitro for binding to the nuclear import receptor karyopherin alpha, which mediates the nuclear import of cellular proteins. A single peptide was identified in which the cis and trans photoisomers bind the receptor differentially. The strategy used to obtain this peptide is systematic and empirical; therefore, it is potentially applicable to any peptide-receptor system.
Park
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SB
Texas A&M University, Department of Chemistry, PO Box 30012, College Station, TX 77842-3012, USA.
Standaert
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RF
D000595:Amino Acid Sequence; D000818:Animals; D001671:Biochemistry / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D019913:Nuclear Localization Signals / Q000737:chemistry / Q000378:metabolism; D009687:Nuclear Proteins / Q000138:chemical synthesis* / Q000235:genetics / Q000378:metabolism*; D056489:Nucleoplasmins; D019151:Peptide Library; D010455:Peptides; D010750:Phosphoproteins / Q000138:chemical synthesis* / Q000235:genetics / Q000378:metabolism*; D010777:Photochemistry / Q000379:methods; D014982:Xenopus laevis / Q000235:genetics; D028901:alpha Karyopherins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D008024:Ligands; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D056489:Nucleoplasmins; C449101:PKL-1c peptide; D019151:Peptide Library; D010455:Peptides; D010750:Phosphoproteins; D028901:alpha Karyopherins