Design of DNA-conjugated polypeptide-based capture probes for the anchoring of proteins to DNA matrices.
A new method for protein surface functionalization was developed that utilizes DNA-conjugated artificial polypeptides to capture recombinant target proteins from the solution phase and direct their deposition onto DNA-functionalized matrices. Protein capture is accomplished through the coiled-coil association of an engineered pair of heterodimeric leucine zippers. Incorporating half of the zipper complex directly into the polypeptides and labeling these polymers with ssDNA enables the polypeptide conjugates to form intermediate linkages that connect the target proteins securely to DNA-functionalized supports. This synthetic route provides an important alternative to conventional DNA-conjugation techniques by allowing proteins to be outfitted site-specifically with ssDNA while minimizing the need for postexpression processing. We demonstrate these attributes by (i) using the capture probes to prepare protein microarrays, (ii) demonstrating control over enzyme activity via deposition of DNA, and, (iii) synthesizing finite-sized, multiprotein complexes that are templated on designed DNA scaffolds in near quantitative yield.
Schweller
Ryan M
RM
Department of Chemistry, Rice University, 6100 Main Street, MS 142 Houston, Texas 77005, USA.
Constantinou
Pamela E
PE
Frankel
Nicholas W
NW
Narayan
Priyanka
P
Diehl
Michael R
MR
D000595:Amino Acid Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D055767:Immobilized Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D015202:Protein Engineering
D004277:DNA, Single-Stranded; D055767:Immobilized Proteins; D010455:Peptides; D004247:DNA