Ribosome-mediated incorporation of peptides and peptidomimetics
AssigneeArizona Board of Regents on behalf of Arizona State University
InventorsSidney HECHT, Larisa (Liza) DEDKOVA, Rumit MAINI, Sandipan ROY CHOWDHURY, Rakesh PAUL
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Abstract
Modified ribosomes that were selected using a dipeptidyl-puromycin aminonucleoside are used to mediate site-specific incorporation of one or more peptides and peptidomimetics into protein in a cell free translation system. In addition, new fluorescent dipeptidomimetics have been synthesized and incorporated into proteins, as well as modified proteins containing one or more non-naturally occurring dipeptides.
Claims11
A genetically modified ribosome having a 23S rRNA sequence modification selected from the group consisting of one or more modification(s) in Table 5.
An in vitro translation system including materials sufficient to effect translation from a mRNA, comprising:
- a genetically modified ribosome having a 23S rRNA sequence modification selected from the group consisting of one or more modification(s) in Table 5.
A method for producing a modified peptide, polypeptide, or protein, comprising the step of:
- providing to an in vitro translation system a tRNA with a modified peptide or peptidomimetic, said system including materials sufficient to effect translation from a mRNA and containing a genetically modified ribosome having a 23S rRNA sequence modification selected from the group consisting of one or more modification(s) in Table 5.
A novel fluorescent dipeptidomimetic selected from the group consisting of dipeptidomimetics 6-13.
A modified protein, said protein containing a dipeptide (analogue) or combination of dipeptide analogues not occurring naturally in said modified protein.
A modified protein containing a non-naturally occurring dipeptide selected from the group consisting of:
- dipeptides containing fluorophores, dipeptides containing dehydroamino acids, dipeptides containing embedded phosphoramide or sulfonamide functionalities, and dipeptide isosteres as shown in Table 6.
A modified protein containing at least one fluorescent dipeptide not including a thiodipeptide and useful for detection and sensing applications.
A modified protein containing at least one fluorescent dipeptide and an additional fluorescent amino acid useful for Förster resonance energy transfer (FRET) measurements, or a quencher useful for photoinduced electron transfer (PET) measurements, to enable the measurement of distances and dynamic changes in protein structure.
Dependencies are inferred from the claim text.