Oligonucleotide-directed self-assembly of proteins: semisynthetic DNA--streptavidin hybrid molecules as connectors for the generation of macroscopic arrays and the construction of supramolecular bioconjugates.
Modified biomolecules were used for the non-covalent assembly of novel bioconjugates. Hybrid molecules were synthesized from short single-stranded DNA and streptavidin by chemical methods using a heterobispecific crosslinker. The covalent attachment of an oligonucleotide moiety to streptavidin provides a specific recognition domain for a complementary nucleic acid sequence, in addition to the four native biotin-binding sites. These bispecific binding capabilities allow the hybrid molecules to serve as versatile connectors in a variety of applications. Bifunctional constructs have been prepared from two complementary hybrid molecules, each previously conjugated to biotinylated immunoglobulin G or alkaline phosphatase. The use of nucleic acid sequences as a template for the formation of an array of proteins is further demonstrated on two size scales. A macroscopic DNA array on a microtiter plate has been transformed into a comparable protein chip. A nano-scale array was made by hybridizing DNA-tagged proteins to specific positions along a RNA or DNA sequence. The generation of supramolecular bioconjugates was shown by quantitative measurements and gel-retardation assays.
Niemeyer
C M
CM
Center for Advanced Biotechnology, Boston University, MA 02215.
Sano
T
T
Smith
C L
CL
Cantor
C R
CR
D001426:Bacterial Proteins / Q000737:chemistry; D001483:Base Sequence; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D012313:RNA / Q000737:chemistry; D019809:Streptavidin
D001426:Bacterial Proteins; D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D011506:Proteins; D012313:RNA; D019809:Streptavidin