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Components for protein nucleic acid conjugation using HhC biocatalysis. (A) Pregnenolone, a bifunctional sterol. (B) Oxime formation: condensation of pregnenolone and alkoxyamines. (C) FRET assay for HhC activity: optical reporter monitors HhC sterylation activity by loss of FRET from precursor, C−H−Y. (D) Steramers: sterylation of benzaldehyde modified nucleic acid using oxime bond chemistry. (E) Separation of steramer from starting oligonucleotide by RP-HPLC.
