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Nucleic Acids Research, 1994, Vol. 22, No. 25 5530-5539
© 1994

Articles

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

Christof M. Niemeyer*, Takeshi Sano, Cassandra L. Smith and Charles R. Cantor

Center for Advanced Biotechnology, Boston University 36 Cummington Street, Boston, MA 02215, USA

*To whom correspondence should be addressed at. University of Bremen, Biotechnology and Molecular Genetics Department (FB 02), Haferwende 12, Postfach 33 04 40, 28334 Bfemen, Germany

Received August 25, 1994. Revised November 9, 1994. Accepted November 18, 1994.

Modified biomolecules were used for the non-covalent assembly of novel bioconjugates. Hybrid molecules were synthesized from short single-stranded DNA and streptavldln by chemical methods using a heterobi-speciflc crossllnker. The covalent attachment of an oligonucleotide moiety to streptavldln provides a specific recognition domain for a complementary nucleic acid sequence, in addition to the four native biotln-blndlng sites. These bispeciflc binding capabilities allow the hybrid molecules to serve as versatile connectors in a variety of applications. Blfunctional constructs have been prepared from two complementary hybrid molecules, each previously conjugated to blotlnylated immunoglobulln 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.


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